Pseudomonas strain h16 and its use

Abstract

The application provides a Pseudomonas strain H16 and application thereof. The Pseudomonas H16 provided by the application has a preservation number of CCTCC NO: M 2023940. The strain is isolated from a part of Ageratina adenophora in Yulin City of Guangxi Zhuang Autonomous Region, has antagonistic effects on various pathogenic bacteria, and has a wide antibacterial spectrum. The strain shows excellent preventive and therapeutic effects on anthracnose of Prunus salicina, and the preventive and therapeutic effects are 91.7% and 79.6% respectively. The application provides a scientific basis for development and utilization of the biocontrol bacteria.

Description

Technical Field

[0001] This invention belongs to the field of agricultural microbial technology and relates to a strain of Pseudomonas monsensis H16 and its application in the control of anthracnose in plum trees. Background Technology

[0002] Anthracnose, caused by Colletotrichum, is one of the major diseases affecting plum trees. This disease can infect plum trees throughout their entire growth period, primarily affecting the leaves, and secondarily the fruit. Severely infected plum trees exhibit premature leaf drop, branch dieback, and fruit rot, significantly impacting the quality and yield of the plums.

[0003] Currently, the control of anthracnose in plum trees mainly relies on chemical control. However, long-term and excessive application of chemical agents can lead to varying degrees of drug resistance in pathogens, and can also cause environmental pollution and food safety issues. Therefore, developing sustainable and green control technologies is particularly important. Biological control offers the advantage of providing a safe and effective method to avoid the drawbacks of chemical control. It not only reduces the environmental pollution risks associated with chemical agents but also prevents pathogens from developing resistance. Furthermore, some biological agents are live microorganisms that can colonize the plant surface and within the plant after application, significantly prolonging their effectiveness and regulating the microecological balance of the host plant. Therefore, developing microbial preparations that can replace chemical agents is an important direction in the research of biological control of plant diseases. Summary of the Invention

[0004] The purpose of this invention is to provide a strain of Pseudomonas monsensis, H16, and its application in the control of anthracnose in plum trees. It has a strong inhibitory effect on anthracnose in plum trees and also has antibacterial activity against a variety of plant pathogens.

[0005] To achieve the objectives of this invention, the inventors isolated and purified a bacterial strain, designated H16, from the flowers of *Age of Hordeum vulgare*, which exhibits antagonistic activity against the pathogen causing anthracnose in plum trees. After culturing on LA medium for 2–3 days, the strain produced milky-white, opaque, round colonies with a moist, viscous, slightly raised surface and neat edges; it was Gram-negative; and exhibited weak salt tolerance, failing to grow at a salt concentration of 7%. The gene sequences of the strain were uploaded to NCBI, with accession numbers 16S rDNA (OQ410272) and gyrB (OQ578988). Based on the combined phylogenetic analysis of the 16S rDNA and gyrB genes and the physiological and biochemical characteristics, the strain was identified as *Pseudomonas monsensis*. The strain was deposited on June 5, 2023, at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M 2023940, located at Wuhan University, Wuhan, China.

[0006] A strain of Pseudomonas monsensis, strain H16, with accession number CCTCCM2023940.

[0007] The application of the aforementioned Pseudomonas strain H16 in the control of plant pathogens.

[0008] The pathogens mentioned are *Botrytis cinerea*, *Dragon fruit canker fungus*, *Pseudomonas violacea*, *Rhizoctonia solani*, *Helicobacter mangois*, *Phytophthora lichrysogenum*, *Fusarium oxysporum*, white rot fungi, *Alternaria mangois*, *Pseudomonas mangois*, *Metacarpa mangois*, *Metacarpa mangois*, *Metacarpa mangois*, *Anthracis mangois*, *Anthracis mangois*, and *Bacillus oryzae*, *Oryza sativa*.

[0009] The plant pathogen is *Anthracnose spp.*

[0010] The prevention and control method involves spraying the fermentation broth of the aforementioned Pseudomonas strain H16 onto the leaf surface of the pearl plum seedlings to control anthracnose in the pearl plum.

[0011] The viable cell count of the fermentation broth is 1×10⁻⁶. 7 CFU / ml.

[0012] This invention provides a strain of Pseudomonas H16 with broad-spectrum antibacterial activity. This bacterium has inhibitory effects on a variety of fruit crop pathogens, including Staphylococcus aureus, Clostridium violaceum, Rhizoctonia solani, Helicobacter mangois, Phytophthora lichee, Fusarium moniliforme, white rot fungi, Alternaria mangois, Epicoccus mangois, Metasporum tobira, Metasporum tobira, Anthracnose tobira, Anthracnose tobira, and Bacillus oryzae of rice.

[0013] The fermentation broth and volatile gases of Pseudomonas H16 provided by this invention can inhibit the growth of anthrax mycelia. The fermentation broth causes the pathogenic mycelia to swell and become vacuolated, turning them black and dark brown; while the volatile substances cause the mycelia to deform and branch.

[0014] The volatile components of Pseudomonas H16 provided by this invention are mainly 14 compounds, including ketones, alkenes, aldehydes, alcohols and benzoquinones, most of which are ketones, with tridecanone having the highest relative content at 14.092%.

[0015] The fermentation broth of this *Pseudomonas* strain H16 showed a 21.2% initial screening inhibitory effect against *Anthracnose spp.* in *Prunus mume*, while the volatile substances showed a 73.1% inhibitory effect against the pathogen. The volatile substances exhibit excellent inhibitory effects, allowing for non-contact control, and are particularly suitable for large-scale, enclosed orchards.

[0016] This invention provides the application of *Pseudomonas* H16 in the control of plum anthracnose. Through greenhouse control trials, *Pseudomonas* H16 exhibited excellent preventative and curative effects, at 91.7% and 79.6%, respectively.

[0017] Preservation information for Pseudomonas monsensis strain H16:

[0018] Strain classification and nomenclature: Pseudomonas monsensis

[0019] Microbial culture preservation number: CCTCC NO: M 2023940

[0020] Depository: China Center for Type Culture Collection, Wuhan University, Wuhan, China

[0021] Deposit date: June 5, 2023 Attached Figure Description

[0022] Figure 1 The fermentation product of strain H16 inhibited the growth of Anthracnose mycelium (A: control CK, B: effect of H16 fermentation product on inhibiting the growth of Anthracnose mycelium).

[0023] Figure 2 The volatile substances of strain H16 inhibit the growth of *Anthracis pilosa* mycelium (A: control CK, B: effect of volatile substances of H16 on inhibiting the growth of *Anthracis pilosa* mycelium).

[0024] Figure 3 The diagram shows the phylogenetic results of strain H16 based on 16S rDNA and gyrB gene. Strain XYAH1 is strain H16, which is Pseudomonas monsensis.

[0025] Figure 4 The images show the morphological characteristics (A) and Gram staining results (B) of strain H16 on LA plates.

[0026] Figure 5 The growth of strain H16 in media with salt contents of 2%, 5%, 7%, and 10% are shown.

[0027] Figure 6 The effect of strain H16 on the mycelial morphology of *Anthracis pilosa* DS1-G-1 was investigated. A represents the control; B represents the effect of the strain's fermentation broth on mycelial morphology; and C represents the effect of the strain's volatile substances on mycelial morphology. Detailed Implementation

[0028] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0029] The following strains are all preserved in the applicant's laboratory and can be distributed externally.

[0030] Example 1: Isolation of Pseudomonas H16

[0031] Healthy *Ageratum conyzoides* plant samples were collected from Yulin City, Guangxi Zhuang Autonomous Region, placed in sealed bags, labeled, and quickly brought back to the laboratory for separation. The flowers of the *Ageratum conyzoides* samples were cut into small pieces using a sterilized scalpel and surface-sterilized in sterile separation cups: immersed in 75% ethanol for 10 seconds, immersed in 2% sodium hypochlorite for 1 minute, and then rinsed three times with sterile water. The samples were then transferred to centrifuge tubes containing an appropriate amount of sterile water and grinding beads, and thoroughly ground. The mixture was then diluted 10-fold, 100-fold, and 100-fold with sterile water, respectively. 100 μL of each dilution was plated on LA medium (10 g tryptone, 5 g yeast extract, 10 g sodium chloride, 15 g agar powder, 1000 mL distilled water) and incubated at 30°C. Single colonies were observed daily for growth; single colonies were streaked onto fresh LA plates for purification. This process was repeated until purified strain H16 was obtained. After purification, single colonies were picked and placed in LB liquid medium (10g tryptone, 5g yeast extract, 10g sodium chloride, 1000mL distilled water) and cultured at 30℃ and 180rpm for 24h with shaking. The shaken bacterial solution was then mixed with sterile 50% glycerol at a ratio of 1:1 in cryovials and stored at -80℃ for later use at -20℃.

[0032] Example 2: Biological characteristics of Pseudomonas H16

[0033] 1. Morphological characteristics on the culture medium

[0034] Use a pipette to draw 20 μL of bacterial suspension of strain H16 and spread it onto an LA plate. After sealing with a sealing film, incubate in a 30°C incubator for 48 hours. Then, remove the culture dish and observe and record the basic characteristics such as colony morphology, color, and texture.

[0035] After being cultured on LA medium for 2–3 days, strain H16 colonies are opaque and off-white, with a moist and viscous surface and regular edges. Figure 4 (A)

[0036] 2. Gram staining

[0037] Heat the inoculation needle with an alcohol lamp until it is red-hot. After cooling, pick a small amount of bacterial growth and spread it onto a clean glass slide in a drop of sterile water. Air dry the slide, then briefly heat it over an alcohol lamp flame to fix the bacteria. After the slide cools, add a mixture of crystal violet and let it stand for 1 minute, then rinse with running water. Induce with iodine solution for 1 minute, rinse with clean water, and blot dry with absorbent paper. Decolorize with 95% ethanol, dripping until the decolorizing solution is colorless (approximately 30 seconds). Stain with safranin for 2–3 minutes, rinse with water, and air dry. Examine under a 100x optical microscope: deep purple indicates Gram-positive bacteria; red indicates Gram-negative bacteria.

[0038] Strain H16 stains red with Gram, indicating it is a Gram-negative bacterium. Figure 4 (B)

[0039] 3. Salt tolerance and salt requirement

[0040] The tested strains were inoculated into LB medium containing different concentrations of NaCl (2%, 5%, 7%, and 10%) and cultured with shaking at 30°C and 180 rpm. Uninoculated LB medium was used as a control. The growth of the strains at different concentrations was observed after 72 hours.

[0041] Strain H16 has poor salt tolerance and cannot grow when the salt content reaches 7%. Figure 5 ).

[0042] Example 3: Molecular biological identification of Pseudomonas H16

[0043] Using DNA from strain H16 as a template, amplification was performed using multiple pairs of primers.

[0044] Universal primers for bacterial 16S rDNA:

[0045] 27F(5'-AGAGTTTGATCCTGGCTCAG-3'),

[0046] 1492R(5'-GGTTACCTTGTTACGACTT-3').

[0047] Bacterial gyrB gene primers:

[0048] UP-1(5'-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3'),

[0049] UP-2r (5'-AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3').

[0050] PCR amplification was performed using a 25 μL reaction system: ddH2O 9.5 μL, 2×Phanta Max Master Mix 12.5 μL, template DNA 1 μL, upstream primer (10 μm) 1 μL, and downstream primer (10 μm) 1 μL.

[0051] Amplification procedure:

[0052] 16S rDNA: pre-denaturation at 95℃ for 5 min, denaturation at 94℃ for 1 min, annealing at 58℃ for 30 s, extension at 72℃ for 1 min, repeated 32 times, followed by extension at 72℃ for 10 min.

[0053] gyrB: Pre-denaturation at 94℃ for 5 min, denaturation at 94℃ for 1 min, annealing at 57℃ for 1 min, extension at 72℃ for 90 s, cycled 40 times, followed by extension at 72℃ for 10 min.

[0054] The amplified PCR products were purified and bidirectionally sequenced by Sangon Biotech (Shanghai) Co., Ltd. The sequences were assembled using DNAMAN 8.0 software, then aligned using BLAST on NCBI. A phylogenetic tree was constructed using MEGA 5.2 and Clustal_X software based on the Neighbor-joining method, and the strain sequences were uploaded to NCBI. The accession numbers for strain H16 are: 16S rDNA (OQ410272); gyrB (OQ578988). The combined phylogenetic tree of 16S rDNA and gyrB genes indicates that strain H16 is *Pseudomonas monsensis*. Figure 3 ).

[0055] Example 4: Physiological and biochemical identification of Pseudomonas H16

[0056] Referring to the methods in Bergey's Manual of Bacterial Identification (8th Edition) and the Manual of Systematic Identification of Common Bacteria, various physiological and biochemical indicators of the strain were determined.

[0057] 1. Citrate utilization test

[0058] Inoculate by streaking on a slant of the culture medium and incubate at the appropriate temperature for 3–7 days. A positive result is indicated by an alkaline culture medium (indicator is pink), otherwise a negative result is indicated.

[0059] 2. Catalase

[0060] The strain was cultured on a plate for 24 hours. A small loop of the strain was then applied to a glass slide containing 3% hydrogen peroxide using a platinum wire inoculation loop. The presence of bubbles indicated a positive result, while the absence of bubbles indicated a negative result.

[0061] 3. Methyl Red (MR)

[0062] Inoculate the test bacteria into the culture medium, with two replicates each time, and incubate at a suitable temperature for 2–6 days (the incubation time can be appropriately extended if the result is negative). Add one drop of methyl red reagent to the culture medium; red indicates a positive methyl red test, and yellow indicates a negative reaction (color change range 4.4 red to 6.0 yellow).

[0063] 4. VP Measurement

[0064] Mix an equal volume of culture medium with 40% sodium hydroxide. Add a small amount of creatine. If the culture medium turns red after 10 minutes, the test is positive. Sometimes, a longer time is required for the red reaction to appear.

[0065] 5. Starch hydrolysis

[0066] Inoculate fresh, activated cultures onto plates and incubate at the appropriate temperature. After 2–5 days of incubation, once distinct colonies have formed, add Lugol's iodine solution (same as Gram staining) to the plate. If the plate turns blue-black and a clear zone remains unchanged around the colonies, it indicates a positive result for starch hydrolysis; otherwise, it remains blue-black and indicates a negative result.

[0067] 6. Cellulose decomposition

[0068] Dispense the basal culture medium into test tubes. Immerse a strip of filter paper in the medium, ensuring part of the paper strip is above the surface of the medium. Inoculate the test tubes with the test bacteria, using uninoculated tubes as blank controls. Incubate at room temperature for 1–4 weeks. A positive result is indicated by the filter paper strip decomposing into a clump of fibers, breaking, or thinning. A negative result is indicated by no change in the filter paper strip.

[0069] 7. Gelatin liquefaction

[0070] Take 18-24 h slant culture for stab inoculation, and prepare two uninoculated blank controls. Incubate at 20℃ for 2-30 days. Observe growth and gelatin liquefaction at room temperature below 20℃. If bacteria have grown, and the gelatin surface is without depressions and forms a stable curd, the gelatin hydrolysis is negative. If the gelatin curd partially or completely becomes a flowable liquid below 20℃, the gelatin hydrolysis is positive.

[0071] 8. Utilization of sugar alcohols

[0072] The tested substrates, including sugar alcohols, were lactose, D(+)-xylose, D-(+)-maltose, inositol, D-mannitol, sucrose, D-(-)-fructose, D-sorbitol, D(+)-glucose, D-(+)-galactose, eugenol, and L(+)-rhamnose. The final concentration was 1%, and the bacteria were inoculated with bacterial suspension. After three consecutive generations, the bacteria that grew were considered positive.

[0073] Physiological and biochemical characteristics of Pseudomonas H16: positive results for citrate utilization, catalase, methyl red, starch hydrolysis, gelatin liquefaction, and cellulose hydrolysis; negative results for VP test; can utilize D-(+)-maltose, inositol, D-mannitol, D-(-)-fructose, sucrose, and D(+)-glucose; but cannot utilize D-sorbitol, D(+)-xylose, lactose, D-(+)-galactose, eugenol, and L(+)-rhamnose.

[0074] Example 5: Antibacterial effect of Pseudomonas H16 against Anthrax rhamnoides DS1-G-1

[0075] 1. Antibacterial effect of Pseudomonas H16 fermentation broth on Anthracnose DS1-G-1

[0076] The test bacteria were cultured in LB medium at 30°C and 200 rpm with shaking for 24 hours. A small amount of bacterial suspension was streaked onto a 9 cm diameter PDA plate using an inoculation loop. Anthrax bacteria were cultured on PDA medium for 5 days. 5 mm diameter bacterial blocks were punched from the edge of the colonies using a punch, and then inoculated 5 mm away from the bacteria with hyphae facing down. Uninoculated treatments served as controls. Each treatment was replicated three times. After 5 days of incubation at 28°C, the colony radius was measured, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [(Control colony radius - Treatment colony radius) / Control colony radius] × 100%

[0077] Antibacterial effect such as Figure 1 As shown, the calculated inhibition rate of strain H16 against anthrax bacteria was 21.2%.

[0078] 2. The antibacterial effect of volatile substances from Pseudomonas H16 on Anthracnose DS1-G-1

[0079] The bacteria to be tested were cultured in LB medium at 30℃ and 200 rpm for 24 h with shaking. 200 μL of bacterial fermentation broth was then spread onto LA plates. Using a 5 mm diameter punch, mycelial cakes were collected from the edge of pathogen colonies cultured for 5-7 days and inoculated onto the center of a PDA plate with the mycelium facing down. The PDA plate inoculated with the pathogen was placed on top, and the LA plate inoculated with the bacterial fermentation broth was placed on the bottom; the two plates were then inverted and sealed. LA plates without bacterial fermentation broth served as a control. Each treatment was repeated in triplicate, and the plates were incubated at 28℃. After 5 days of incubation at 28℃, the colony diameter was measured, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [(Control colony diameter - Treatment colony diameter) / (Control colony diameter - Mycelial cake diameter)] × 100

[0080] Antibacterial effect such as Figure 2 As shown, the calculated inhibition rate of strain H16 against *Anthracnose bacillus* of *Prunus cerevisiae* was 73.1%.

[0081] 3. After conducting the above-mentioned plate confrontation and plate-on-plate tests, a small amount of mycelium was picked and its growth status was observed under a microscope. The results are as follows: Figure 6 As shown in the image. Under an optical microscope, the control group colonies exhibited dense, robust hyphae at the edges with few branches and a regular growth direction. After the plate confrontation test, the hyphae turned black and dark brown, and the microscope clearly showed that the hyphae morphology deviated from the normal shape, becoming swollen and vacuolated; while the volatile substances of the biocontrol bacteria caused the hyphae to deform and branch, but no swelling was observed.

[0082] Example 6: Control Experiment of Pseudomonas H16 on Plum Anthracnose

[0083] Under greenhouse conditions, an indoor control experiment was conducted on strain H16 using two-year-old pearl plum seedlings. (1) Preventive treatment: Healthy pearl plum seedlings were selected, and the leaves were inoculated with a biocontrol bacterial suspension (10 g / L) by spraying. 7 CFU / mL) was sprayed until the bacterial solution condensed into water droplets on the leaves but did not drip. A 0.05% Tween 20 solution was used as a control instead of the bacterial solution. After culturing in bags at 28℃ for 24 hours, a suspension of pathogenic spores (1×10⁻⁶ CFU / mL) was sprayed. 6 (1 spores / mL), continue bagging and moisturizing culture. Inoculate 3 pearl plum seedlings, each seedling inoculated with 15 leaves. After inoculation with pathogen, observe regularly, keep moisturized, record the disease situation after 5 days, and calculate the incidence rate, severity of disease, disease index and greenhouse control effect. (2) Treatment: First spray pathogen spore suspension, then spray biocontrol bacterial suspension, the inoculation method is the same as the above disease prevention test.

[0084] Disease severity is expressed as the percentage of lesion area to the total leaf surface area, and is divided into 6 levels: 0, no disease; 1, lesion area ≤ 5%; 2, lesion area 6-15%; 3, lesion area 16-25%; 4, lesion area 26-50%; 50% or more. The formulas for calculating the incidence rate, disease index, and greenhouse control effectiveness are as follows:

[0085] Incidence rate (%) = 100 × (number of diseased leaves / total number of leaves)

[0086] Disease index = [∑(number of leaves at each disease level × representative value for that level) / (total number of leaves × 9)] × 100

[0087] Greenhouse control efficacy (%) = (CK disease index) - (T disease index) / (CK disease index) × 100

[0088] The results of the greenhouse control experiment are shown in Table 1. The biocontrol strain H16 has excellent preventive and curative effects, at 91.7% and 79.6%, respectively.

[0089] Table 1 Results of greenhouse pest control experiment

[0090]

[0091] Example 7: Identification of volatile components of Pseudomonas H16

[0092] Methods: The test strain was cultured in LB liquid medium at 30℃ and 180 rpm for 24 h with shaking. Then, 200 μL of the test bacterial solution was transferred to a 100 mL headspace vial containing 20 mL of LA solid medium. After drying in a clean bench, the vial was sealed with a silicone rubber septum and incubated at 30℃ for 3 days. A sealed headspace vial without bacterial solution served as a control. After equilibration in a 60℃ water bath for 5 min, a pre-aged extraction tip (50 / 30 μm DVB / CAR / PDMS, American) was inserted into the headspace vial for 50 min. The extraction tip was then removed from the vial and immediately inserted into the gas chromatograph injection port. Desorption was performed at 280℃ for 5 min, followed by gas chromatography / mass spectrometry (GC / MS, 7890A-5975CMSD, American). The mass spectra of the obtained gas components were compared with the international standard database (Library of the National Institute of Standards and Technology 2017, NIST2017), and combined with time retention index (RI) and AMDIS spectrum analysis to determine the chemical composition of each unknown compound. Then, the compounds present in the control were removed from the bacterial volatile components.

[0093] The operating conditions for gas chromatography / mass spectrometry are as follows: 1) GC conditions: Column: DB-5ms quartz capillary column (30m×0.25mm×0.25μm); Temperature program: Initial temperature 40℃, hold for 5 min, increase to 120℃ at 4℃ / min, hold for 3 min, then increase to 220℃ at 10℃ / min, hold for 10 min; then increase to 280℃ at 30℃ / min and hold for 10 min. Carrier gas (He) flow rate is 1.0 mL / min; Injection mode: splitless injection. 2) MS conditions: Electron impact (EI) ionization source, electron energy 70 eV; ion source temperature 250℃; quadrupole temperature 150℃; auxiliary heating temperature 250℃; mass scan range m / z 35~450.

[0094] The volatile components of Pseudomonas H16 are mainly 14 compounds, including ketones, alkenes, aldehydes, alcohols and benzoquinones. The majority of these are ketones, with tridecanone having the highest relative content at 14.092% (Table 3).

[0095] Table 3 Volatile Substances

[0096]

[0097] Example 8: Antibacterial spectrum of Pseudomonas H16

[0098] Indicator bacteria (plant pathogenic fungi) preserved at 4℃ were aseptically transferred onto PDA plates and activated at 28℃ for later use. Test bacteria were cultured in LB medium at 30℃ and 200 rpm with shaking for 24 hours. A small amount of bacterial suspension was streaked 2 cm from the edge of a 9 cm diameter PDA plate using an inoculation loop. Anthracnose bacteria were cultured on PDA medium for 5 days. 5 mm diameter bacterial blocks were punched at the edge of the colonies using a punch, and then inoculated 5 mm from the bacteria with hyphae facing down. Uninoculated treatments served as controls, and each treatment was replicated in triplicate. After 5 days of incubation at 28℃, the colony radius was measured, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [(Control colony radius - Treatment colony radius) / Control colony radius] × 100%.

[0099] Strain H16 showed inhibitory effects against all 14 tested pathogens, with the best inhibitory effect against *Pseudomonas mangifera* (58.8%). The inhibition rate against *Anthracis pilosa* was 20.4% (Table 4), comparable to the initial screening result of 21.2%. Figure 1 ).

[0100] Table 4 Antibacterial spectrum

[0101] TEY11A-1 Botryosphaeria dothidea 19.9±1.2c HF Neoscytalidium dimidiatum, the pathogen that causes canker in dragon fruit 0.0±0.0d EGY16A-1 Pseudofusicoccum violaceum 18.1±0.6cd YMWK Rhizoctonia solani 20.7±1.2bc YB33-1 Exserohilum sp. 31.5±10.9abc LZ1-3-2 Peronophythora litchii 14.3±1.1d TEY15-1 Fusarium sulawesiense 32.6±6.7b FY27-1 White rot fungus Phanerochaete sordida 18.5±7.3abc YB43-2 Alternaria brassicae 49.6±19.7a YB53-2 Epicoccum sorghinum 58.8±17.3a BY58-2 Diaporthe phoenicicola 15.2±0.8c YB3 Diaporthe phaseolorum 25.0±9.5c DS1-G-1 Colletotrichum fructicola, anthrax fungus of the pearl plum 20.4±22.3a GZ15-1 Mango anthrax fungus Colletotrichum fructicola 31.7±11.3b 6-6 rice blast fungus Magnaporthe oryzae 21.3±2.1c

Claims

1. A plant Pseudomonas monsensis Strain H16, with accession number CCTCC NO: M 2023940.

2. The application of strain H16 according to claim 1 in the control of plant pathogens, wherein the plant pathogens are *Botrytis cinerea*, *Pseudomonas violacea*, *Rhizoctonia solani*, *Helicobacter mangois*, *Phytophthora lichrysogenum*, *Fusarium oxysporum*, white rot fungi, *Alternaria mangois*, *Pseudomonas mangois*, *Metacarpa mangois*, *Metacarpa mangois*, *Anthracnose fungus of *Pseudomonas ... and *Bacillus oryzae*, the causal agent of rice blast.

3. The application according to claim 2, wherein the plant pathogen is *Anthracnose fungus of *Prunus cerasifera*.

4. The application according to claim 3, wherein the prevention and control involves spraying the fermentation broth of strain H16 according to claim 1 onto the leaf surface of the pearl plum seedlings to achieve the prevention and control of anthracnose fungus in pearl plum.

5. In the application according to claim 4, the viable count of the fermentation broth is 1 × 10⁻⁶. 7 CFU / ml.

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