Bacillus amyloliquefaciens and application thereof

Abstract

The invention discloses a bacillus amyloliquefaciens strain and application thereof. The bacillus amyloliquefaciens SF-00595 disclosed by the invention is obtained by carrying out heat resistance domestication on bacillus amyloliquefaciens SDF-005, the preservation number of the bacillus amyloliquefaciens SF-00595 is CCTCC (China Center For Type Culture Collection) NO: M 2025830, and the bacillus amyloliquefaciens SF-00595 can be used as a biopesticide for preventing and treating papaya anthracnose, fig anthracnose and the like. The bacillus amyloliquefaciens SF-00595 strain is subjected to heat resistance domestication, so that the death rate of the strain during spray drying of fermentation liquor is reduced, and the content of viable bacteria in bacterial powder is increased.

Description

Technical Field

[0001] The present invention belongs to the technical field of microorganisms, and in particular relates to a Bacillus amyloliquefaciens strain and application thereof. Background Art

[0002] Fig( Ficusc arica Linn.) is a perennial subtropical deciduous fruit tree, small tree or shrub. Figs have high nutritional value. In addition to nutrients such as sugar, protein, fat and minerals, the fruit also contains trace elements such as copper, iron, zinc, manganese, selenium and rich vitamins. Both the fruit and leaves of figs have certain oxidizing properties. The ethanol extract has a significant broad-spectrum antibacterial effect, and the latex hexane extract has a strong bactericidal effect. Fig latex extract can inhibit the formation and growth of capillaries in umbilical vein endothelial cells. Eating figs can effectively prevent cancer and other chronic diseases, and can also lower blood sugar and blood lipids. It is known as the "guardian of human health in the 21st century." Papaya ( Pseudocydonia sinensis (Thouin) CK Schneid.) is a shrub or small tree in the genus Chaenomeles in the Rosaceae family. Papaya has both economic and medicinal value, primarily for its medicinal uses. Fresh medicinal papaya fruit has attracted considerable attention for its unique nutritional profile.

[0003] Figs and papayas have become emerging economic forestry industries. However, due to climate change, shifts in cultivation practices, and the rapid expansion of cultivated areas, fig and papaya pests and diseases have become increasingly frequent and recurring. Anthracnose, in particular, has become a serious threat to the healthy development of the industry. Currently, fig and papaya pest and disease control primarily relies on chemical pesticides, resulting in increasing pathogen resistance, decreased control effectiveness, and excessive pesticide residues, seriously impacting both fruit quality and ecological safety.

[0004] Microbial-derived biopesticides have become a research hotspot in the field of biopesticides and the main choice for biopesticide registration due to their advantages such as fast reproduction speed, long-lasting control effect, multiple development and utilization channels, low resistance, simple fermentation process and low production cost. Bacillus amyloliquefaciens ) is an effective biocontrol bacterium and a stable biopesticide strain developed in recent years. Bacillus amyloliquefaciens SDF-005 was isolated from rhizospheric soil of a walnut forest in the southern mountainous area of Jinan City, Shandong Province, using the dilution separation method. This strain is deposited under the CGMCC No. 6672. This strain has a strong inhibitory effect against Colletotrichum walnutii and is used to control walnut anthracnose. Based on this, it is necessary to screen for new strains and expand their applications. Summary of the Invention

[0005] In view of the above problems, the present invention provides a Bacillus amyloliquefaciens strain and its application. After domestication of the heat resistance of Bacillus amyloliquefaciens SDF-005 (CGMCC No. 6672), a strain of Bacillus amyloliquefaciens SF-00595 with strong heat resistance and capable of preventing and controlling papaya anthracnose and fig anthracnose is obtained. This strain can be used as a biological pesticide to prevent and control papaya anthracnose, fig anthracnose, etc., and has good application prospects.

[0006] The above object of the present invention is achieved by the following technical solutions: The Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ), SF-00595, is obtained by domestication of the heat resistance of Bacillus amyloliquefaciens SDF-005 (CGMCC No. 6672) (capable of withstanding high temperature of 95 °C), and has been deposited at the China Center for Type Culture Collection (CCTCC) on April 21, 2025. The deposit address is: Wuhan University, Wuhan, China. The deposit number is CCTCC NO: M2025830. It has the following microbiological characteristics: The cells are rod-shaped, Gram-positive bacteria, with peritrichous flagella, facultative anaerobes. The colonies cultured on NA medium are round, white, flat, with a rough surface, a circular protrusion in the middle, and neat edges. A pellicle is formed in liquid culture, and the liquid color is brownish-yellow, turbid, with precipitation. The above strain is stored in NA medium for ordinary culture and stored in 20% glycerol for long-term storage.

[0007] The present invention also provides a biological bactericide with Bacillus amyloliquefaciens SF-00595 as the active ingredient.

[0008] The preparation method of the above biological bactericide is as follows: 1) Shaking flask culture: Pick a small amount of colonies of Bacillus amyloliquefaciens SF-00595 from the test tube slant and transfer them to the seed medium, and culture them on a shaker at 28 - 32 °C for 12 - 14 h; 2) Seed culture: Add the shaker-cultured bacterial liquid to a seed tank filled with the seed medium for culture. The culture temperature is 35 - 40 °C, the initial pH is 7.0 - 7.5, and the culture time is 12 - 14 h to obtain the seed liquid; 3) Fermentation culture: Transfer the seed liquid to the fermentation medium for culture to obtain the fermentation broth. The culture temperature is 35 - 40 °C, the initial pH is 7.0 - 7.5, the inoculation amount (mass ratio) is 3 - 10%, and the culture time is 20 - 30 h. Then the fermentation broth is further made into a biological bactericide.

[0009] The seed medium is (weight percentage): soybean meal powder 3.2%, corn starch 2.2%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, pH 7.0 - 7.5.

[0010] The fermentation medium is (by weight percentage): soybean meal powder 5.9%, corn starch 4.8%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, with a pH of 7.0 - 7.5.

[0011] Furthermore, the above fermentation broth is spray-dried with a water-soluble carrier. The spray-dried bacterial powder is poured into a mixer, and the water-soluble carrier is continuously added and stirred evenly to obtain a biological bacterial agent.

[0012] The present invention also provides the use of Bacillus amyloliquefaciens SF-00595 and its biological bacterial agent for controlling (preventing and treating) papaya anthracnose.

[0013] The present invention also provides the use of Bacillus amyloliquefaciens SF-00595 and its biological bacterial agent for controlling fig anthracnose.

[0014] The present invention also provides a method for controlling papaya anthracnose and fig anthracnose, which includes applying the biological bacterial agent of Bacillus amyloliquefaciens SF-00595 to papayas and figs with anthracnose.

[0015] The technical effects of the present invention are as follows: 1. The Bacillus amyloliquefaciens SF-00595 strain has been domesticated for heat resistance, reducing the mortality rate of the strain during spray drying of the fermentation broth and increasing the viable bacteria content in the bacterial powder.

[0016] 2. By using the spraying method, compared with the water control, the preventive effects of the biological bacterial agent of Bacillus amyloliquefaciens SF-00595 at treatment 2 (100 and 200 times dilution) on papaya anthracnose and fig anthracnose both reach over 70%, and the treatment effects of the biological bacterial agent of Bacillus amyloliquefaciens SF-00595 at treatment 3 (100, 200, and 300 times dilution) are over 60%, showing good preventive and treatment effects.

[0017] 3. Since the biological bacterial agent of Bacillus amyloliquefaciens SF-00595 has no drug residues, no toxic and side effects, and can reduce the use of chemical pesticides, it has good application prospects in biological control of papaya anthracnose and fig anthracnose. Description of the Drawings

[0018] Figure 1 It is a single colony morphology diagram of Bacillus amyloliquefaciens SF-00595; Figure 2 It is the antibacterial effect of Bacillus amyloliquefaciens SF-00595 against fig anthracnose bacteria. Among them, Figure A shows the antibacterial effect of Bacillus amyloliquefaciens SF-00595 against fig anthracnose bacteria; Figure B is the control inoculated only with fig anthracnose bacteria; Figure 3To study the antibacterial effect of Bacillus amyloliquefaciens SF-00595 against Colletotrichum gloeosporioides, Figure A shows the antibacterial effect of Bacillus amyloliquefaciens SF-00595 against Colletotrichum gloeosporioides on papaya fruits; Figure B shows the control inoculated only with Colletotrichum gloeosporioides. Detailed implementation mode

[0019] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that without departing from the spirit and scope of the present invention, the details and forms of the technical solutions of the present invention can be modified or replaced, but these modifications and replacements all fall within the protection scope of the present invention.

[0020] Example 1: Isolation and identification of Bacillus amyloliquefaciens SF-00595 1. Isolation: The new strain of Bacillus amyloliquefaciens of the present invention is obtained by domestication of the heat resistance of Bacillus amyloliquefaciens SDF-005 (CGMCC No. 6672). The acquisition method is as follows: (1) Test strain: Bacillus amyloliquefaciens SDF-005 (CGMCC No. 6672); (2) Preparation of culture medium plates: Melt the potato dextrose agar medium (PDA), pour it into sterile petri dishes respectively, and cool to make PDA plates; the formula of the PDA medium is: 200 g of potatoes (peeled and cut into pieces), 20 g of glucose, 14 g of agar, and 1000 mL of distilled water; (3) Use an inoculation loop to pick up well-grown colonies of Bacillus amyloliquefaciens SDF-005 into NB culture solution, and shake-culture at 26 °C and 160 r / min for 36 h. Use a pipette to suck 1 mL of the bacterial solution, dissolve it in 9 mL of sterile water, shake well, and place it in a water bath at 60 °C, 70 °C, 80 °C, 85 °C, 90 °C, and 95 °C for 30 min respectively. Each temperature is a treatment, and each treatment is repeated 3 times. After cooling to room temperature (about 25 °C), use the dilution separation method to purify and obtain single colony strains. The strains obtained under different temperature treatments are numbered SDF-00560, SDF-00570, SDF-00580, SDF-00585, SDF-00590, and SF-00595 in turn.

[0021] (4) Using Colletotrichum gloeosporioides on fig fruits as the target, the antibacterial activity was measured by the in vitro confrontation culture method. The strain ability was evaluated by comprehensively considering the heat resistance ability and antibacterial effect of the strains, and 1 strain with strong inhibitory activity against Colletotrichum gloeosporioides on fig fruits was screened, numbered SF-00595.

[0022] 2. Strain Identification (1)Microbiological characteristics: The cells are rod-shaped, Gram-positive bacteria, with peritrichous flagella and facultative anaerobes. The colonies cultured on NA medium are circular, white, flat, with a rough surface, a circular protrusion in the middle, and a neat edge (see Figure 1 ). A pellicle is formed in liquid culture, and the liquid color is brownish-yellow, turbid, with precipitation.

[0023] (2)Molecular biological characteristics: The sequencing result of the 16S rRNA gene sequence of the above SF-00595 strain is shown in SEQ No.1; gyrB The sequencing result of the gene sequence is shown in SEQ No.2.

[0024] The 16S rRNA amplified sequence of the SF-00595 strain and the sequence of 1 housekeeping gene ( gyrB )were aligned in the NCBI database. The homology of the 16S rRNA sequence with Bacillus amyloliquefaciens LT212 (CP000560) is 99.86%, gyrB The homology of the gene with Bacillus amyloliquefaciens GXU-1 is 100%. Combining with the morphological characteristics of the SF-00595 strain, it is determined that SF-00595 belongs to Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ).

[0025] This strain was deposited in the China Center for Type Culture Collection (abbreviated as CCTCC) on April 21, 2025. Address: China. Wuhan. Wuhan University, China Center for Type Culture Collection, Deposit number: CCTCC NO: M 2025830.

[0026] Example 2 Verification of the indoor effect of Bacillus amyloliquefaciens SF-00595 against Colletotrichum gloeosporioides and Colletotrichum caricicola 1. Experimental method Using the plate confrontation culture method, the test Colletotrichum gloeosporioides ( Colletotrichum aenigma ), Colletotrichum caricicola ( Colletotrichum fioriniae ) mycelial cakes (5 mm) were transferred to the center of the plate, and the Bacillus amyloliquefaciens SF-00595 strain was symmetrically inoculated 2 cm away from the mycelial cake. It was cultured in a biochemical incubator at 28 °C, and each treatment was repeated 3 times. When the mycelium of the control group (only inoculating a 5 mm Colletotrichum mycelial cake in the center of the plate) covered the plate, the cross-cross method was used to measure the colony diameter and the diameter of the inhibition zone, and the average value and relative inhibition rate were calculated.

[0027] 2. Results From Table 1 and Figure 2 , Figure 3It can be seen that Bacillus amyloliquefaciens SF-00595 has a strong inhibitory effect on the mycelial growth of Colletotrichum gloeosporioides and Colletotrichum caricicola. The inhibition zones are 12.6 mm and 11.9 mm respectively, and the mycelial growth inhibition rates are 81.5% and 80.0% respectively.

[0028] Table 1 Determination of indoor antibacterial activity of Bacillus amyloliquefaciens SF-00595 against Colletotrichum gloeosporioides and Colletotrichum caricicola

[0029] Example 3 Preparation of Bacillus amyloliquefaciens SF-00595 bacterial agent The seed liquid medium is (weight percentage): soybean meal powder 3.2%, corn starch 2.2%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, pH 7.0 - 7.5.

[0030] The fermentation medium is (weight percentage): soybean meal powder 5.9%, corn starch 4.8%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, pH 7.0 - 7.5.

[0031] The preparation method includes the following steps: 1) Shaking flask culture: Pick a small amount of colonies of Bacillus amyloliquefaciens SF-00595 from the test tube slant and transfer them to the seed liquid medium, and culture them on a shaker at 30 °C for 12 - 14 h; 2) Seed liquid culture: Add the bacterium solution cultured by shaking to a seed tank (250 L) filled with the seed liquid medium for culture. The culture temperature is 37 °C, the initial pH is 7.0 - 7.5, the ventilation volume is 15 m , , ,

[0031] ,

[0032] , , , , 3 , , 3 , , / h, and the culture time is 12 - 14 h; 3) Fermentation culture: Transfer the seed liquid to the fermentation medium for culture. The culture temperature is 37 °C, the initial pH is 7.0 - 7.5, the liquid loading volume is 70%, the inoculation amount is 10%, the culture time is 24 h, and the ventilation volume is 220 m 3 / h. Using this medium for fermentation culture, the number of bacteria can reach 100 billion / g; 4) Dosage of auxiliary materials: Add water-soluble carrier industrial glucose according to 10% of the mass of the fermentation broth, and stir evenly in the batching tank; 5) Spray drying: The inlet air temperature is 170 °C, and the outlet air temperature is 75 - 70 °C; 6) Compound: Pour the spray-dried bacterial powder into a mixer, add water-soluble carrier industrial glucose, and stir well for 30 min to make it evenly mixed, and prepare a bacterial agent with a viable bacteria content of 60 billion / g.

[0032] Example 4 Verification of the effect of Bacillus amyloliquefaciens SF-00595 bacterial agent on Colletotrichum gloeosporioides of fig This example provides experiments related to the activity of Bacillus amyloliquefaciens SF-00595 against fig anthracnose.

[0033] 1. Methods 1.1 Test agent: Bacillus amyloliquefaciens SF-00595 bacterial agent (active bacterial content 60 billion / g).

[0034] 1.2 Test crops and control targets: The test crop was fig (Green Early) and the target disease was anthracnose. Fig leaves of uniform size and health, unexposed to the pesticide, were collected from the experimental base. They were rinsed three times with sterile water, air-dried, and placed in a fresh-keeping box (16 cm × 10 cm × 6 cm) containing soaked filter paper until ready for use.

[0035] 1.3 Methods 1.3.1 Determination of preventive effect Using a hand sprayer (E532), Bacillus amyloliquefaciens SF-00595 inoculum (100-fold, 200-fold, and 300-fold dilutions) was sprayed on the surface of fig leaves to form a uniform film. Fig leaves sprayed with water served as a control. 24 h after treatment, a sterilized inoculation needle was used to slightly puncture the surface of the fig leaves to form a microwound. 100 μL of anthracnose spore suspension (10 6 Spores / mL) were dropped on the wound. 15 leaves were treated for each time, and each leaf was inoculated at one place. The inoculated leaves were placed in an incubator (25℃; L﹕D = 12 h﹕12 h; relative humidity above 85%) for incubation and the disease development was observed. After 7 days of incubation, the diameter of the lesions was measured and the average value was taken to calculate the prevention effect. The experiment was repeated 4 times.

[0036] 1.3.2 Determination of treatment efficacy Fig leaves collected for the experiment were first inoculated with an anthracnose spore suspension according to the above method. 24 hours later, the leaves were sprayed with the same concentrations (100x, 200x, and 300x dilutions) as described above. Control leaves were sprayed with water. Each treatment consisted of 15 leaves, with one inoculation site per leaf. The experimental figs were incubated in an incubator (25°C; L / D = 12 h / 12 h; relative humidity above 85%) and observed for disease development. After 7 days of incubation, the diameter of the lesions was measured and the average value was used to calculate the control efficacy. The experiment was repeated four times.

[0037] 1.4 Statistics Control effect (%) = (control lesion diameter - treatment lesion diameter) / control lesion diameter × 100.

[0038] 2. Results 2.1 Preventive effect of Bacillus amyloliquefaciens SF-00595 agent on anthracnose of Ficus carica The results in Table 2 show that the Bacillus amyloliquefaciens SF-00595 agent has a good preventive effect on anthracnose of Ficus carica, and the preventive effects at 100-fold, 200-fold, and 300-fold dilutions are 78.95%, 73.49%, and 62.57% respectively.

[0039] Table 2 Preventive effect of Bacillus amyloliquefaciens SF-00595 agent on anthracnose of Ficus carica

[0040] 2.2 Therapeutic effect of Bacillus amyloliquefaciens SF-00595 agent on anthracnose of Ficus carica The results in Table 3 show that the Bacillus amyloliquefaciens SF-00595 agent has a good therapeutic effect on anthracnose of Ficus carica, and the therapeutic effects at 100-fold, 200-fold, and 300-fold dilutions are 68.84%, 65.96%, and 64.40% respectively.

[0041] Table 3 Therapeutic effect of Bacillus amyloliquefaciens SF-00595 agent on anthracnose of Ficus carica

[0042] 3. Summary The Bacillus amyloliquefaciens SF-00595 agent has a good control effect on anthracnose of Ficus carica. The preventive effects of the 100-fold and 200-fold dilutions are greater than their therapeutic effects, and the preventive effect increases with the increase of the agent concentration. The preventive effect of the 300-fold dilution is equivalent to the therapeutic effect. After dilution at 100-fold and 200-fold, the preventive effects can reach 78.95% and 73.49%, and the therapeutic effects reach 68.84% and 65.96%. The three treatments have good preventive and therapeutic effects on anthracnose of Ficus carica and can effectively control the harm of anthracnose of Ficus carica.

[0043] Example 5: Verification of the effect of Bacillus amyloliquefaciens SF-00595 agent on anthracnose of Chaenomeles sinensis This example provides relevant experiments on the Bacillus amyloliquefaciens SF-00595 agent against anthracnose of Chaenomeles sinensis.

[0044] 1. Method 1.1 Test agent: Bacillus amyloliquefaciens SF-00595 agent (effective bacteria content: 60 billion / g).

[0045] 1.2 Test crops and control objects: Test crops: Chaenomeles sinensis; Control object: Anthracnose.

[0046] Collect papaya fruits of the same size and health status that have not been exposed to pesticides in the experimental base. After rinsing 3 times with sterile water and drying, place them in a beaker (1L) containing wet filter paper for later use.

[0047] 1.3 Test methods 1.3.1 Determination of preventive effect Use a manual sprayer (E532) to spray the formulated Bacillus amyloliquefaciens SF-00595 bactericide at different concentrations (100-fold, 200-fold, 300-fold dilutions) on the surface of papaya fruits to form a uniform drug film, with papaya fruits sprayed with clear water as the control. At 24 h after treatment, slightly prick the surface of the papaya fruits with a sterilized inoculation needle to form micro-wounds, and drop 100 μL of the anthracnose spore suspension (10 6 spores / mL) on the wounds. Use 10 fruits for each treatment, inoculate one place on each fruit, place the inoculated fruits in a sterile beaker (1L) (25 °C; L:D = 12 h:12 h; relative humidity above 85%) for cultivation and observe the disease incidence. After 7 days of cultivation, measure the diameter of the disease spots and take the average value to calculate the control effect. The experiment was repeated 4 times.

[0048] 1.3.2 Determination of treatment effect The papaya fruits collected in the experiment were first inoculated with the anthracnose spore suspension according to the above method, and after 24 h, the liquid medicine of the same concentration (100-fold, 200-fold, 300-fold dilutions) was sprayed according to the above method, and the control leaves were sprayed with clear water. Use 10 fruits for each treatment, inoculate one place on each fruit, place the experimental papayas in a sterile beaker (1L) (25 °C; L:D = 12 h:12 h; relative humidity above 85%) for cultivation and observe the disease incidence. After 7 days of cultivation, measure the diameter of the disease spots and take the average value to calculate the control effect. The experiment was repeated 4 times.

[0049] 1.4 Data statistics Same as Example 4.

[0050] 2. Results 2.1 Preventive effect of Bacillus amyloliquefaciens SF-00595 bactericide on papaya anthracnose Table 4 shows that the Bacillus amyloliquefaciens SF-00595 bactericide has a good preventive effect on papaya anthracnose, and the preventive effects at 100-fold, 200-fold, and 300-fold are 77.72%, 75.02%, and 72.45% respectively.

[0051] Table 4 Preventive effect of Bacillus amyloliquefaciens SF-00595 bactericide on papaya anthracnose

[0052] 2.2 Therapeutic effect of Bacillus amyloliquefaciens SF-00595 agent on papaya anthracnose Table 5 shows that the Bacillus amyloliquefaciens SF-00595 agent has a good therapeutic effect on papaya anthracnose, and the therapeutic effects at 100-fold, 200-fold, and 300-fold are 68.64%, 66.05%, and 63.34% respectively.

[0053] Table 5 Therapeutic effect of Bacillus amyloliquefaciens SF-00595 agent on papaya anthracnose

[0054] 3. Summary The preventive effects of the three treatments of the Bacillus amyloliquefaciens SF-00595 agent are greater than its therapeutic effects, and the effects increase with the increase of the agent concentration. The three treatments all have good preventive effects (the preventive effects of all three treatments reach more than 70%) and therapeutic effects (the therapeutic effects of all three treatments reach more than 60%) on papaya anthracnose, and can effectively control the harm of papaya anthracnose.

Claims

1. Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) SF-00595, the preservation number of the strain is CCTCC NO: M 2025830.

2. The Bacillus amyloliquefaciens SF-00595 according to claim 1, characterized in that, The strain is resistant to high temperature of 95°C.

3. A biological bactericide with the Bacillus amyloliquefaciens SF-00595 described in claim 1 as the active ingredient.

4. The preparation method of the biological bacterial agent according to claim 3, characterized in that, It includes the following steps: 1) Shaking culture: Pick colonies of Bacillus amyloliquefaciens SF-00595 from the test tube slant and transfer them to the seed liquid medium, and culture them on a shaker at 28-32°C for 12-14 h with shaking; 2) Seed liquid culture: Add the bacterium liquid cultured with shaking to the seed tank filled with the seed liquid medium for culture. The culture temperature is 35-40°C, the initial pH is 7.0-7.5, and the culture time is 12-14 h to obtain the seed liquid; 3) Fermentation culture: Transfer the seed liquid to the fermentation medium for culture to obtain the fermentation broth. The culture temperature is 35-40°C, the initial pH is 7.0-7.5, the inoculation amount is 3-10%, and the culture time is 20-30 h. Then, the fermentation broth is further made into a biological bactericide.

5. The preparation method of the biological bacterial agent according to claim 4, characterized in that, The seed liquid medium, by weight percentage, is: soybean meal powder 3.2%, corn starch 2.2%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, with a pH of 7.0-7.

5.

6. The preparation method of the biological bacterium agent according to claim 4, characterized in that, The fermentation medium, by weight percentage, is: soybean meal powder 5.9%, corn starch 4.8%, glucose 0.5%, manganese sulfate 0.02%, and the rest is water, with a pH of 7.0-7.

5.

7. The preparation method of the biological bacterial agent according to claim 4, characterized in that, Add the fermentation broth to a water-soluble carrier for spray drying, pour the spray-dried bacterial powder into a mixer, and continue to add the water-soluble carrier and stir evenly to obtain the biological bactericide.

8. Use of the Bacillus amyloliquefaciens SF-00595 described in claim 1 or the biological bactericide described in claim 3 for preventing and treating papaya anthracnose.

9. Use of the Bacillus amyloliquefaciens SF-00595 described in claim 1 or the biological bactericide described in claim 3 for preventing and treating fig anthracnose.

10. A method for preventing and controlling anthracnose of papaya or anthracnose of fig, characterized in that, The method includes applying the Bacillus amyloliquefaciens SF-00595 biological bactericide described in claim 3 to papayas or figs with anthracnose.

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