Paenibacillus polymyxa FangSG106 as well as application, product and method thereof

Abstract

The invention relates to paenibacillus polymyxa FangSG106 as well as application, a product and a method thereof, and belongs to the technical field of microorganisms. The invention provides paenibacillus polymyxa FangSG106, the preservation number of the paenibacillus polymyxa FangSG106 is CGMCC (China General Microbiological Culture Collection Center) No.31655. The invention also provides application of the paenibacillus polymyxa FangSG106 with the preservation number of CGMCC No.31655 in the aspects of non-treatment-destination bacteriostasis, plant disease prevention and treatment and plant growth promotion, and based on the paenibacillus polymyxa FangSG106, the application of the paenibacillus polymyxa FangSG106 in the aspects of non-treatment-destination bacteriostasis, plant disease prevention and treatment and plant growth promotion is The invention provides a plant growth promoter, a biopesticide, a method for preventing and treating plant diseases, a method for promoting plant growth and a bacteriostasis method. The paenibacillus polymyxa FangSG106 disclosed by the invention has broad-spectrum antibacterial activity, and can be used for effectively preventing and treating plant diseases and promoting plant growth.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to Paenibacillus polymyxa Fang SG106 and applications, products and methods thereof. Background Art

[0002] In crop production, plant diseases are one of the important factors leading to yield reduction. Cucumber is an important economic crop. Cucumber yield and quality are seriously damaged by cucumber wilt and damping-off caused by Fusarium oxysporum and Rhizoctonia solani. These two pathogenic fungi are soil-borne diseases with wide geographical distribution, strong destructive effects and difficult to control, which greatly restricts the development of cucumber planting industry and leads to significant agricultural and economic losses. Therefore, research on the prevention and control of these two pathogens is of great significance.

[0003] Chemical pesticides are currently the most common method for plant disease control. This method is effective and highly operable, but it can cause ecological pollution, make pathogens resistant, and ultimately threaten human health. Compared with chemical pesticides, biological control is an eco-friendly, sustainable and safe method for plant disease control. The use of antagonistic microorganisms to replace traditional chemical pesticides has huge market application prospects. Biological control mainly promotes the growth and development of host plants to form systemic resistance by means of antagonistic bacteria secreting antibiotics and lipopeptide compounds, increasing the dissolution of inorganic nutrients, and inducing host resistance. In addition, bacteria reproduce relatively quickly, and multiple biological control mechanisms often work at the same time, making it difficult for pathogens to develop resistance.

[0004] Bacteria from various genera such as Bacillus, Paenibacillus, Pseudomonas, and Rhizobium have been identified as biological control agents. Paenibacillus polymyxa belongs to the genus Paenibacillus of the Bacillaceae family. It is a Gram-positive, facultative anaerobic, neutrophilic, spore-bearing rod-shaped bacterium with a cell wall that does not contain endotoxins, is harmless to humans and animals, and is widely distributed in nature. Many strains of Paenibacillus polymyxa have broad-spectrum antibacterial activity. The bacteria can secrete a variety of antimicrobial active substances, such as cell wall degrading enzymes, polymyxins, fusarium, and polypeptide antibiotics, which can directly kill plant pathogens by destroying the cell wall of plant pathogens or causing abnormal development of the hyphae of plant pathogenic fungi. In addition, Paenibacillus polymyxa has a variety of growth-promoting mechanisms, including the production of indoleacetic acid, the secretion of siderophores to dissolve mineral phosphates, and the induction of plant resistance.

[0005] In summary, Paenibacillus polymyxa is an ideal bacterium for use as a biofertilizer and biocontrol agent, and has great potential for development and application.

[0006] However, the number of strains of Paenibacillus polymyxa that can be used in practice is limited, and more new strains of Paenibacillus polymyxa need to be developed. At the same time, in practice, the agricultural microbial environment is complex, and some harmful microorganisms or competing microorganisms can affect the colonization and growth of Paenibacillus polymyxa. Therefore, the field needs to develop new strains of Paenibacillus polymyxa that have both broad-spectrum antibacterial properties and biological control capabilities. Summary of the invention

[0007] In order to solve the technical problems in the prior art that the number of Bacillus polymyxa that can be actually applied is limited and there is a lack of new strains of Bacillus polymyxa that have broad-spectrum antibacterial properties and biological control capabilities, the present invention provides a Bacillus polymyxa FangSG106 with broad-spectrum antibacterial properties and its applications, products and methods.

[0008] The technical solution of the present invention is as follows:

[0009] A Paenibacillus polymyxa FangSG106, characterized in that its deposit number is CGMCC No.31655.

[0010] The Paenibacillus polymyxa FangSG106 with the deposit number of CGMCC No.31655 is used in inhibiting bacteria in non-therapeutic destinations, preventing and controlling plant diseases, and promoting plant growth.

[0011] The antibacterial spectrum includes: Rhizoctonia solani, Magnaporthe oryzae, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium oxysporum cucumber transformation, Fusarium wilt, Botrytis cinerea, Aeromonas verticillatus, Aeromonas jantaensis, Aeromonas punctatus, Vibrio splenici, and Vibrio anguillarum;

[0012] Preferably, the prevention of plant diseases refers to: preventing and controlling cucumber wilt and cucumber damping-off;

[0013] Preferably, the promoting plant growth refers to: promoting the growth of root length, plant height, stem thickness, fresh weight and dry weight of cucumber seedlings.

[0014] A biological pesticide comprises a pharmacologically active ingredient, wherein the pharmacologically active ingredient comprises: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655.

[0015] The biological pesticide further comprises: pharmaceutically acceptable excipients.

[0016] A plant growth promoter comprises active ingredients, wherein the active ingredients include: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655.

[0017] The plant growth promoter also includes auxiliary materials.

[0018] The invention discloses a method for preventing and controlling plant diseases, which adopts Paenibacillus polymyxa FangSG106 with the preservation number of CGMCC No.31655 to prevent and control plant diseases.

[0019] A method for promoting plant growth adopts Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655 to promote plant growth.

[0020] The invention discloses a bacteriostatic method, which adopts Paenibacillus polymyxa FangSG106 with the preservation number of CGMCC No.31655 to inhibit bacteria.

[0021] The beneficial effects of the present invention are as follows:

[0022] Compared with the prior art, the present invention has the following outstanding significant advantages: the polymyxa bacillus FangSG106 provided by the present invention has a broad-spectrum antibacterial property. The antibacterial zone experiment proves that the polymyxa bacillus FangSG106 has an antibacterial effect on Rhizoctonia solani, Magnaporthe oryzae, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium oxysporum cucumber transformation, Cabbage wilt, Botrytis cinerea, Aeromonas verticillatus, Aeromonas jenda, Aeromonas punctata, Vibrio splendens, and Vibrio anguillarum. At the same time, it is confirmed by plant experiments that the polymyxa bacillus FangSG106 can effectively prevent and treat cucumber wilt and cucumber damping-off, and promote the growth of cucumber seedlings including root length, plant height, stem thickness, fresh weight, dry weight, etc., and significantly promote plant growth compared with the control group. The polymyxa bacillus FangSG106 has a fast spore production speed and a high spore rate, and shows great application prospects in the prevention and control of agricultural fungal diseases.

[0023] The deposit information of Paenibacillus polymyxa FangSG106 of the present invention is as follows:

[0024] Deposit number: CGMCC No.31655

[0025] Storage date: August 12, 2024

[0026] Classification name: Paenibacillus polymyxa

[0027] Depository: China National Microbiological Culture Collection Administration General Microbiology Center

[0028] Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a curve diagram of the spore formation rate of the five strains of Paenibacillus polymyxa with the fastest spore production rate in Experimental Example 2 of the present invention.

[0030] Figure 2 The colony morphology (a), bacterial morphology (b) and spore morphology (c) of Paenibacillus polymyxa FangSG106 of Experimental Example 2 of the present invention.

[0031] Figure 3 This is the inhibitory effect of Paenibacillus polymyxa FangSG106 of Experimental Example 2 of the present invention on seven pathogenic fungal strains: (a) Rhizoctonia solani, (b) Magnaporthe oryzae, (c) Sclerotinia sclerotiorum, (d) Fusarium oxysporum, (e) Fusarium oxysporum cucumber transformed type, (f) Fusarium wilt, and (g) Botrytis cinerea. The upper figure is the CK group and the lower figure is the strain FangSG106 group.

[0032] Figure 4 This is a qualitative result diagram of the plate production of protease (a), amylase (b), cellulase (c) and peroxidase (d) by Paenibacillus polymyxa FangSG106 in Experimental Example 3 of the present invention.

[0033] Figure 5 This is a qualitative plate diagram of the ability of Paenibacillus polymyxa FangSG106 to dissolve inorganic phosphorus in Experimental Example 4 of the present invention.

[0034] Figure 6 This is a graph showing the experimental results of Paenibacillus polymyxa FangSG106 promoting the growth of potted cucumber seedlings in Experimental Example 6 of the present invention, wherein the 3 seedlings on the left are the control group, and the 3 seedlings on the right are the treatment group. DETAILED DESCRIPTION

[0035] The present invention is further described in detail below with reference to specific embodiments, experimental examples and drawings, but the embodiments of the present invention are not limited thereto.

[0036] 1. Sources of biological materials

[0037] (i) The pathogenic fungi used in Experimental Example 2 of the present invention: Rhizoctonia solani, Sclerotinia sclerotiorum and Fusarium oxysporum f. sp. conglutinans were kindly donated by the National Key Laboratory of Microbiology, Huazhong Agricultural University.

[0038] Pathogenic fungi: Magnaporthe oryzae and Fusarium oxysporum were kindly donated by Beijing Academy of Agriculture and Forestry Sciences.

[0039] Pathogenic fungi: Fusarium oxysporum cucumber transformation type ACCC39326 (Fusarium oxysporum) and Botrytis cinerea ACCC37347 (Botrytis cinerea) were purchased from China Agricultural Microbiological Culture Collection Center.

[0040] Pathogenic bacteria: Pseudomonas syringae, Agrobacterium tumefaciens, Acidovorax citrulli and Xanthomonas citrisubsp. citri were isolated and obtained in our laboratory in the early stage. Clostridioides difficile ATCC43593 was purchased from Guangdong Microbial Culture Collection Center. Serratia marcescens was donated by the Institute of Agricultural Resources and Agricultural Zoning, Chinese Academy of Agricultural Sciences.

[0041] (ii) Aeromonas veronii, Aeromonas jandaei, Aeromonas punctata, Vibriosplendidus and Vibrio anguillarum of Experimental Example 9 of the present invention were isolated and obtained from diseased fish tissues or water environment samples in the early stage of this laboratory.

[0042] Aeromonas veronii was isolated and obtained by the applicant's laboratory from diseased fish tissues or water environment samples in the early stage. It is the Aeromonas veronii recorded in the article "Chen F, Sun J, Han Z, et al. Isolation, Identification and Characteristics of Aeromonas veronii From Diseased Crucian Carp (Carassiusauratus gibelio). Front Microbiol, 2019, 10: 2742." The applicant promises to release it to the public within 20 years from the date of application of the present invention for the purpose of verifying the technical effect of the present invention.

[0043] Aeromonas jandaei was isolated and obtained by the applicant's laboratory from diseased fish tissues or water environment samples in the early stage. It is the Aeromonas jandaei recorded in the article "Chen M, Xue M, Chen J, et al. Isolation, identification and characterization of Aeromonas jandaei from diseased Chinese soft-shell turtles. J Fish Dis. 2024, 47(4): e13919." The applicant promises to release it to the public within 20 years from the date of application of the present invention for the purpose of verifying the technical effect of the present invention.

[0044] Aeromonas punctata was isolated by the applicant's laboratory from diseased fish tissues or water environment samples in the early stage. It is the Aeromonas punctata recorded in the article "Yingna, Wang Yuan, Qin Bo, et al. Isolation, identification and drug sensitivity test of pathogenic bacteria of discus fish enteritis [J]. Fisheries Science and Technology Information, 2020, 47(05): 245-248." The applicant promises to release it to the public within 20 years from the date of application of this invention to verify the technical effect of the invention.

[0045] Vibrio splendidus was isolated and obtained by the applicant's laboratory from diseased fish tissues or water environment samples in the early stage. It is the Vibrio splendidus recorded in the article "Ningning Liu, Shanshan Zhang, Weiwei Zhang, et al. Vibrio sp. 33a potential bacterial antagonist of Vibrio splendidus pathogenic to sea cucumber (Apostichopus japonicus). Aquaculture, 2017, 470: 68-73." The applicant promises to release it to the public within 20 years from the date of application of the present invention for the purpose of verifying the technical effect of the present invention.

[0046] Vibrio anguillarum was isolated and obtained by the applicant's laboratory from diseased fish tissues or water environment samples in the early stage. It is the Vibrio anguillarum recorded in the article "Yang N, Sun C. The Inhibition and Resistance Mechanisms of Actinonin, Isolated from Marine Streptomyces sp. NHF165, against Vibrio anguillarum. Front Microbiol. 2016, 13; 7: 1467." The applicant promises to release it to the public within 20 years from the date of application of this invention for the purpose of verifying the technical effect of the invention.

[0047] 2. Experimental culture medium

[0048] PDA (Potato Dextrose Agar) agar medium: 200 g peeled potatoes, 20 g glucose, 15 g agar, 1000 mL distilled water, high temperature sterilization at 121°C for 20 min.

[0049] PDB (Potato Dextrose Broth) broth culture medium: 200 g peeled potatoes, 20 g glucose, 1000 mL distilled water, high temperature sterilization at 121°C for 20 min.

[0050] LB (Luria Broth) agar medium: 10 g tryptone, 5 g NaCl, 5 g yeast powder, 15 g agar powder, 1 L distilled water, pH 7.2, autoclave at 121 °C for 15 min.

[0051] LB liquid culture medium: 10 g tryptone, 5 g NaCl, 5 g yeast powder, 1 L distilled water, pH 7.2, autoclave at 121 °C for 15 min.

[0052] BHI medium was purchased from Qingdao Haibo Biotechnology Co., Ltd.

[0053] Protease culture medium: 15 g skim milk powder, 2 g yeast powder, 10 g NaCl, 20 g agar, sterilize at 115 °C for 20 min, pH 7.0.

[0054] Amylase medium: 5g yeast powder, 10g peptone, 5g NaCl, 18g agar, 10g starch, sterilized at 121℃ for 20min, pH 7.0.

[0055] Cellulase detection plate: 2-10 g sodium carboxymethyl cellulose, 1.5 g potassium dihydrogen phosphate, 2.5 g peptone, 0.5 g yeast powder, 2.5 g disodium hydrogen phosphate, 1 L water, 15 g agar, pH 7.0, sterilize at 121°C for 20 min.

[0056] Aniline blue medium: 20 g glucose, 10 g yeast, 0.1 g aniline blue, 1 L water, 15 g agar, pH 7.0, sterilize at 121°C for 20 min.

[0057] Inorganic phosphorus qualitative culture medium: glucose 10g, ammonium sulfate 0.5g, NaCl 0.2g, KCl 0.2g, MgSO4·7H2O0.1g, MnSO4·7H2O 0.005g, FeSO4·7H2O 0.005g, yeast powder 0.5g, Ca3(PO4) 5.0g, distilled water 1L, agar 18g, pH 7.0-7.2.

[0058] The first group of examples, the polymyxa bacillus FangSG106 of the present invention

[0059] This group of embodiments provides a Paenibacillus polymyxa FangSG106, whose deposit number is CGMCC No.31655.

[0060] Any act of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, or promising to sell a strain of Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC NO.31655, and / or using a strain of Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC NO.31655 in combination with other probiotics, and / or using a strain of Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC NO.31655 to antagonize various pathogens, and / or preparing antibacterial products, and / or preparing drugs for improving constipation or regulating intestinal flora, and / or preparing health foods with laxative function or intestinal flora regulating function all fall within the protection scope of the present invention.

[0061] The other probiotics include but are not limited to: Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii lactic acid subsp., Lactobacillus helveticus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Saccharomyces cerevisiae, Delb's spore-forming torula, Candida, Wickham's yeast, Pichia, Saccharomyces boulardii, Torulopsis spp., Saccharomyces cerevisiae, Rhodotorula, Schizosaccharomyces pombe , Saccharomyces cerevisiae, Bacillus thuringiensis, Bacillus laterosporus, Bacillus megaterium, Bacillus colloidal, Bacillus azotobacterium, Bacillus sphaericus, Clostridium butyricum, Bifidobacterium adolescentis, Bifidobacterium angularis, Bifidobacterium animalis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium bovis, Bifidobacterium breve, Bifidobacterium dentate, Bifidobacterium infantis (i.e., Bifidobacterium longum infantis subsp. infantis), Bifidobacterium lactis (i.e., Bifidobacterium animalis subsp. cremoris), Bifidobacterium longum, Bifidobacterium pseudocatenulatum, as well as Bifidobacterium thermophilum and Bifidobacterium acidothermophilum.

[0062] Those skilled in the art can make routine selections or adjustments to pharmaceutical excipients according to actual production needs, combined with conventional technical means or basic common sense of pharmaceutical production processes (for example, "Encyclopedia of Preparation Technology", "Pharmaceutical Preparation Technology", etc.), and then make a strain of Bacillus polymyxa FangSG106 with a preservation number of CGMCC NO.31655 into products with different dosage forms, different storage conditions, and different shelf lives. This is possible and easy to do without technical barriers for those skilled in the art.

[0063] The second group of examples, use of the polymyxa bacillus FangSG106 of the present invention

[0064] This group of embodiments provides the use of Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No. 31655 in inhibiting bacteria in non-therapeutic destinations, preventing and controlling plant diseases, and promoting plant growth.

[0065] In a specific embodiment, the antibacterial spectrum includes: Rhizoctonia solani, Magnaporthe oryzae, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium oxysporum cucumber transformation, Fusarium wilt, Botrytis cinerea, Aeromonas verticillatus, Aeromonas japonicus, Aeromonas punctatus, Vibrio splenici, and Vibrio anguillarum;

[0066] Preferably, the prevention of plant diseases refers to: preventing and controlling cucumber wilt and cucumber damping-off;

[0067] Preferably, the promoting plant growth refers to: promoting the growth of root length, plant height, stem thickness, fresh weight and dry weight of cucumber seedlings.

[0068] The third group of embodiments, the biological pesticide of the present invention

[0069] This group of embodiments provides a biological pesticide. All embodiments of this group have the following common features: the biological pesticide includes a pharmacologically active ingredient, and the pharmacologically active ingredient includes: Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655.

[0070] In a further embodiment, the biological pesticide further comprises: pharmaceutically acceptable excipients.

[0071] In a more specific embodiment, the pharmaceutically acceptable excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0072] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned pharmaceutically acceptable excipients, and prepare the polymyxa bacillus FangSG106 with a deposit number of CGMCC NO.31655 into different dosage forms, such as powders, tablets, injections, oral liquids, suppositories, gels, patches, sprays, granules, etc.

[0073] In a specific embodiment, the dosage form of the drug is selected from: one or more of powder, tablet, liquid, and capsule.

[0074] The fourth group of embodiments, the plant growth promoter of the present invention

[0075] This group of embodiments provides a plant growth promoter. All embodiments of this group have the following common features: the plant growth promoter comprises an active ingredient, and the active ingredient comprises: Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655.

[0076] In a further embodiment, the plant growth promoter further includes: auxiliary materials.

[0077] In a more specific embodiment, the excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0078] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the polymyxa bacillus FangSG106 with a preservation number of CGMCC NO.31655 into different dosage forms, such as powders, tablets, injections, oral liquids, suppositories, gels, patches, sprays, lotions, granules, etc.

[0079] In a specific embodiment, the dosage form of the product is selected from: one or more of powder, tablet, liquid, and capsule.

[0080] The fifth group of embodiments, the method for preventing and controlling plant diseases of the present invention

[0081] This group of embodiments provides a method for preventing and controlling plant diseases. All embodiments in this group have the following common features: Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655 is used to prevent and control plant diseases.

[0082] In a specific embodiment, the plant disease is selected from: cucumber wilt and cucumber damping-off.

[0083] The sixth group of embodiments, the method for promoting plant growth of the present invention

[0084] This group of embodiments provides a method for promoting plant growth. All embodiments in this group have the following common features: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655 is used to promote plant growth.

[0085] In a specific embodiment, the promoting plant growth refers to: promoting the growth of root length, plant height, stem thickness, fresh weight and dry weight of cucumber seedlings.

[0086] Group 7 Embodiments, Method for Promoting Plant Growth of the Present Invention

[0087] This group of embodiments provides a method for inhibiting bacteria. All embodiments in this group have the following common features: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655 is used for inhibiting bacteria.

[0088] In a further embodiment, the antibacterial agent refers to: Paenibacillus polymyxa FangSG106 inhibits Rhizoctonia solani, Magnaporthe grisea, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium oxysporum cucumber transformed type, Fusarium wilt, Botrytis cinerea, Aeromonas verticillaster, Aeromonas natans, Aeromonas punctata, Vibrio splenici, and Vibrio anguillarum.

[0089] Experimental Example 1: Isolation, identification and preservation of Paenibacillus polymyxa

[0090] The sampling sites in this experimental case were the soils from farm vegetable gardens in different urban areas of Hubei Province.

[0091] The specific separation and purification steps are as follows:

[0092] (1) Weigh 5 g of soil sample and add it to 45 ml of sterile water. Incubate it in a constant temperature shaker at 30 °C and 200 rpm for 30 min. After standing for 20 min, obtain a sample suspension.

[0093] (2) 1.8 mL of the soil suspension supernatant obtained in step (1) was transferred to a 2 mL cryovial and treated in an 80°C water bath for 10 min;

[0094] (3) Take 100 μL of the sample suspension obtained in step (2) after water bath, and dilute it in a 10-fold gradient to obtain 10 -1 , 10 -2 , 10 -3 times the sample dilution;

[0095] (4) Take the 10 obtained in step (3) respectively -2 , 10 -3 100 μL of the sample dilution was spread on the LB solid culture medium plate, and the spread LB solid culture medium plate was inverted and cultured in a 30° C. incubator for 48 h; after the culture, colonies grew on the LB solid culture medium plate.

[0096] (5) Pick a single colony grown on the LB solid culture medium plate in step (4), streak it onto a new LB solid culture medium plate using the three-zone streak method, and culture it in a constant temperature incubator at 30° C. for 48 h; observe that only one type of colony morphology is found on the LB solid culture medium plate after culture, and pure colonies are obtained, indicating that the separation and purification is complete.

[0097] (6) The monoclonal culture strain purified in step (5) was cultured in liquid, the cells were collected, the genomic DNA was extracted, the 16SrDNA fragment was amplified using the universal primers 27F and 1492R described in paragraph 58 of Chinese invention patent ZL202210478937.4, the PCR amplification product was detected by agarose gel electrophoresis, and the PCR product was sequenced. The PCR reaction system: 10×buffer 10μL, 10mM dNTP 2μL, 1μL of each upper and lower primer, 2μL of DNA template, 0.5μL of Taq enzyme, 34μL of ddH2O. Pre-denaturation at 95℃ for 10min; then 94℃ for 30s, 60℃ for 30s, 72℃ for 1min for a total of 35 cycles, and then extended at 72℃ for 5min. The PCR product was detected by gel electrophoresis and sent to Wuhan Jinkairui Biotechnology Co., Ltd. for sequencing. The identified gene sequence was compared in the NCBI database using the BLAST tool.

[0098] After PCR molecular identification, a total of 200 strains of Bacillus polymyxa were obtained, and the strains were preserved in ultra-low temperature glycerol tubes in the Micro-Kang Probiotics Strain Resource Bank.

[0099] Experimental Example 2: Primary screening of antagonistic ability of Paenibacillus polymyxa and secondary screening of broad-spectrum antibacterial ability

[0100] (1) The plate confrontation method was used to determine the antagonistic effect and perform the initial screening of strains. Starting from the glycerol tube, 200 strains of Paenibacillus polymyxa were activated on LB solid culture medium plates and cultured in a constant temperature incubator at 30°C for 48 hours to grow single colonies. Four different Paenibacillus polymyxa colonies were taken from each plate and inoculated on a PDA solid plate. Then, the activated Rhizoctonia solani and Fusarium oxysporum cucumber transformation pathogenic fungi plates were taken out, and a round bacterial cake was punched out with a 5 mm diameter puncher and transferred to the center of the PDA culture medium plate. The distance between each indicator fungus and each Paenibacillus polymyxa was equal, and the plate without Paenibacillus polymyxa was used as the blank control. The inoculated plate was inverted and cultured in a 28°C incubator. After the blank control was fully grown on the plate, the results were observed. The presence of an inhibition zone was observed, the strain number was recorded, and the size of the inhibition zone was measured. The 14 strains with the highest inhibition rate were screened from the 200 strains, and further antagonistic strains were screened.

[0101] Bacterial inhibition rate (%) = (control pathogen diameter - treated pathogen diameter) / control pathogen diameter x 100%

[0102] (2) According to the screening results of step (1) of Experimental Example 2, 14 strains of Paenibacillus polymyxa were selected for broad-spectrum antibacterial ability screening using the plate confrontation method, and the experiment was repeated three times. Among them, Paenibacillus polymyxa FangSG106 had the strongest broad-spectrum antibacterial ability, as shown in Tables 1 and Figure 1 As shown, the inhibition rate of the fungus against seven pathogenic fungi was higher than 60%, and the inhibition rates against Rhizoctonia solani, Sclerotinia sclerotiorum, Magnaporthe grisea, Fusarium oxysporum, Fusarium oxysporum cucumber transformed type, Fusarium wilt and Botrytis cinerea were 67.53±1.12%, 62.08±1.35%, 70.13±1.33%, 70.29±1.67%, 72.66±0.53%, 71.15±0.67% and 66.66±0.33%, respectively.

[0103] Table 1. Antibacterial results of FangSG106 strain against seven pathogenic fungi

[0104]

[0105] (3) In addition, the Oxford cup method was used to detect the ability of the polymyxa bacillus FangSG106 strain to inhibit six pathogenic bacteria. The antibacterial experimental method is as follows:

[0106] Preparation of suspensions of four plant pathogenic bacteria: Pseudomonas syringae, Agrobacterium tumefaciens, Acidovorax citrinum and Xanthomonas citri var. citri were inoculated into LB liquid medium at a 2% inoculum, respectively, and cultured in a shaking incubator at 30°C and 220 rpm for 24 h. The bacterial suspension concentration was adjusted to 108 CFU / mL is reserved.

[0107] Preparation of Clostridium difficile suspension: C. difficile was inoculated into a BHI blood plate containing 5% defibrinated sheep blood and cultured at 37°C for 16-20 hours. The concentration of the bacterial suspension was adjusted to 10 8 CFU / mL.

[0108] Preparation of Serratia marcescens suspension: Serratia marcescens was inoculated into LB liquid medium at a 2% inoculum, and cultured at 37°C, 250 r / min, and the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0109] Cool the LB solid medium (BHI solid medium for Clostridium difficile) to about 50°C and mix the pathogenic bacteria into the medium in a certain proportion so that the number of viable pathogenic bacteria is around 10 6 CFU / mL order of magnitude, and then quickly poured into a plate with an Oxford cup placed in advance. After the culture medium cooled and solidified, the Oxford cup was taken out, 150 μL of Paenibacillus polymyxa FangSG106 fermentation liquid was added to the well, and the plates were placed in a constant temperature incubator at the corresponding temperature for 24-48 hours, and then the diameter of the inhibition zone was measured.

[0110] The antibacterial results are shown in Table 2. Paenibacillus polymyxa FangSG106 has a certain antibacterial ability against the six pathogenic bacteria.

[0111] Table 2. Antibacterial results of Paenibacillus polymyxa FangSG106 against six pathogenic bacteria

[0112]

[0113] (4) The 14 strains of Bacillus polymyxa selected from the initial screening of step (1) of Experimental Example 2 were inoculated into solid culture medium and placed in a constant temperature incubator at 30°C for 48 hours for activation. The activated single colonies were picked out and inoculated into shake flasks filled with seed culture medium for cultivation. The shake flasks were filled with 30 mL / 250 mL of liquid and cultured at 30°C and 220 r / min for 8 hours to obtain seed liquid; the seed liquid was transferred to a shake flask filled with optimized culture medium at an inoculum size of 4%, and the shake flasks were filled with 30 mL / 250 mL of liquid and cultured at 30°C and 220 r / min. Sampling was performed for microscopic examination to observe spore formation. The experiment was repeated three times as a reference indicator for strain rescreening.

[0114] The optimized culture medium formula is as follows: 8g glucose, 6g corn syrup powder, 0.3g K2HPO4, 0.5g MgSO4 7H2O, 3g CaCO3, 0.3g NaCl, 0.3g K2SO4, 0.75g yeast powder, 0.75g ammonium sulfate, 1L distilled water, pH 7.0-7.2, sterilized at 121°C for 20min for use.

[0115] The results of the antibacterial and spore-forming ability experiments of 200 bacterial strains were comprehensively compared, and a strain FangSG106 with fast spore-forming speed, high spore-forming rate and significant antagonistic effect was screened out. The number of viable bacteria could reach 3.6×10 after 24 hours of culture. 8 CFU / mL, the spore formation rate can reach more than 90%, and its spore rate and spore production speed are significantly higher than other strains. The strain was sent for preservation, and its preservation information is as follows:

[0116] Deposit number: CGMCC No.31655

[0117] Storage date: August 12, 2024

[0118] Classification name: Paenibacillus polymyxa

[0119] Depository: China National Microbiological Culture Collection Administration General Microbiology Center

[0120] Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0121] Figure 2 This is a curve of spore formation rate of the five strains of Paenibacillus polymyxa that produce spores the fastest.

[0122] Through LB plate culture, the colony color was observed to be light yellow or white, sticky, moist and smooth on the surface, and irregular and light on the edge. Through microscopic observation, the strain's body was rod-shaped, could produce oval spores, and was Gram-positive. The colony morphology (a), bacterial morphology (b), and spore morphology (c) of FangSG106 strain are shown in Figure 1. Figure 3 shown.

[0123] Experimental Example 3: Determination of enzyme production capacity of Paenibacillus polymyxa FangSG106

[0124] Hydrolases have the function of hydrolyzing pathogen cell walls and poisoning pathogens. The main components of fungal cell walls are cellulose, glucan, chitin, followed by proteins and lipids. The ability to produce degradative enzymes is an important characteristic of biocontrol bacteria in inhibiting plant pathogens.

[0125] (1) Detection of the protease production ability of FangSG106 strain

[0126] Use a toothpick to pick up a small amount of FangSG106 colony and spot it on the protease culture medium. Incubate it in a constant temperature incubator at 30℃ for 4 days. Set up 3 replicates and observe whether a transparent circle appears. The larger the transparent circle, the stronger the ability to produce protease. Figure 4 As shown in (a), the FangSG106 strain can produce an obvious transparent zone with a diameter of up to 37.83±2.02 mm, and has a strong ability to produce protease.

[0127] (2) Detection of the ability of FangSG106 strain to produce amylase

[0128] Use a toothpick to pick up a small amount of FangSG106 colony and inoculate it on the amylase culture medium. Incubate it in a 30℃ constant temperature incubator for 4 days, set up 3 replicates, then cover and stain it with dilute iodine solution, rinse it with water, and observe whether a transparent circle appears. The larger the transparent circle, the stronger the ability to produce amylase. The results are as follows Figure 4 As shown in (b), the FangSG106 strain can produce an obvious transparent zone with a diameter of up to 48.67±0.58mm, and has a good ability to produce amylase.

[0129] (3) Detection of the ability of FangSG106 strain to produce cellulase

[0130] Use a toothpick to pick up a small amount of FangSG106 colony and spot it on the cellulase culture medium. Incubate it in a constant temperature incubator at 30℃ for 4 days, set up 3 replicates, add 1g / L Congo red solution to the cultured plate for staining for 1h, then soak and wash it twice with 1M NaCl solution for decolorization, soaking for 30min each time, observe whether there is a cellulose hydrolysis circle around the colony. The larger the hydrolysis circle, the stronger the ability to produce cellulase. The results are as follows Figure 4 As shown in (c), the FangSG106 strain can produce an obvious transparent zone with a diameter of up to 38.16±0.76 mm, and has a good ability to produce cellulase.

[0131] (4) Detection of the ability of FangSG106 strain to produce peroxidase

[0132] Use a toothpick to pick up a small amount of FangSG106 colonies and spot them on the aniline blue enzyme-producing qualitative plate, and culture them in a 30℃ constant temperature incubator for 4 days, set up 3 replicates, and observe whether there is a fading circle. Figure 4 As shown in (d), the plate qualitative method detected that the aniline blue around the FangSG106 bacteria had completely faded, indicating that it had the ability to produce peroxidase.

[0133] The results of enzyme production experiments showed that FangSG106 strain has the ability to synthesize proteases and cellulases. Studies have reported that these activities contribute to plant pathogen antagonism and biological control of plant diseases. In addition, the strain has the ability to produce peroxidase, which has the potential to be used in the treatment of waste crop straw in the field and to improve feed utilization.

[0134] Experimental Example 4: Determination of phosphate solubilization ability of FangSG106 strain

[0135] Use a toothpick to pick up a small amount of FangSG106 colonies and spot them on an inorganic phosphorus-positive plate. Incubate in a 30°C constant temperature incubator for 7 days, set up 3 replicates, and observe whether a transparent circle appears. Figure 5 As shown, a transparent halo appeared around the colony of Paenibacillus polymyxa FangSG106, indicating that the strain has the ability to solubilize phosphate and has the potential to promote plant growth.

[0136] Experimental Example 5: Cucumber Seed Growth Promotion Experiment

[0137] (1) Fermentation broth preparation: Paenibacillus polymyxa FangSG106 strain was inoculated into a 500 mL Erlenmeyer flask containing 100 mL of LB broth at a ratio of 2%, and the flask was cultured in a shaking incubator at 30°C and 220 rpm for 36 h. The viable bacterial count of the fermentation broth was adjusted to 1 × 10 8 CFU / mL is reserved.

[0138] (2) Seed germination: Refer to GB / T 3543.4-1995 Inspection Procedure for Crop Seeds (Germination Test), select plump cucumber seeds and soak them in 75% alcohol for 30 seconds, then rinse with sterile water until there is no alcohol residue; then soak them in 1% sodium hypochlorite solution for 20 minutes, and then rinse with sterile water several times to complete seed disinfection. Place a sterile filter paper in a culture dish (9 cm in diameter), sprinkle the sterilized cucumber seeds (25 seeds / dish) evenly on the filter paper in a clean bench, add sterile water (control group) and FangSG106 strain fermentation liquid that has been cultured for 36 hours (treatment group) two different treatment solutions, keep the filter paper moist, and repeat 3 times for each group. Place in a 30℃ constant temperature incubator to moisturize and germinate, and count the seed germination after 3 days.

[0139] Seed germination rate (%) = number of germinated seeds / number of sown seeds × 100

[0140] Table 3. Effect of FangSG106 fermentation liquid on cucumber seed germination

[0141]

[0142] Note: The data in the table are mean ± standard error. Lowercase letters indicate significant differences in germination rate between the control group and the treatment group (P<0.05).

[0143] As shown in Table 3, when cucumber seeds were treated with the 36h fermentation liquid of FangSG106 strain, the germination rate was 95.33±0.63%, while the germination rate of seeds in the sterile water control group was 86.33±1.67%. There was a significant difference between the groups, indicating that FangSG106 strain can promote seed germination.

[0144] Experimental Example 6: Determination of the ability of FangSG106 strain to promote the growth of cucumber seedlings

[0145] The cucumber seeds with uniform germination and growth in Experimental Example 5 were selected and transferred to sterilized nutrient soil (containing a certain proportion of peat soil, vermiculite, perlite, etc.) potting pots (diameter 18.5 cm, height 16.5 cm, bottom diameter 13 cm). One seedling was inoculated in each pot, and 20 mL of 1.0×10 8 CFU / mL The 36h fermentation broth prepared according to step (1) of Experimental Example 5 was used as the treatment group, and an equal amount of sterile water was inoculated as the blank control group, and then the surface was covered with 1-2 cm of sterilized nutrient soil.

[0146] The sterile water control group and FangSG106 fermentation liquid treatment group used 12 pots of cucumber seedlings, respectively. After the cucumber seedlings were cultured in a greenhouse with a temperature of 25±1℃, a relative humidity of 55%±5%, and a photoperiod of 14 hours of light / 10 hours of darkness for 21 days, the root length, plant height, plant stem diameter, plant fresh weight, and plant dry weight of the plants were measured to evaluate the growth-promoting effect of FangSG106 strain on cucumber. Some pictures of the results of the potted growth-promoting experiment are shown in the figure. Figure 6 shown.

[0147] Table 4. Effect of FangSG106 fermentation liquid on cucumber growth (21 days)

[0148]

[0149] Note: The data in the table are mean ± standard error. Lowercase letters in the same column indicate significant differences in seedling growth between the control group and the treatment group (P<0.05).

[0150] The results of the greenhouse potted test data are shown in Table 4. The root length, stem diameter, plant height, fresh weight and dry weight of cucumber seedlings in the soil of the FangSG106 fermentation liquid treatment group increased by 33.31%, 15.75%, 25.60%, 20.09% and 40% respectively compared with the sterile water control group, indicating that the FangSG106 strain can significantly promote the growth of cucumber plants.

[0151] Experimental Example 7: Experiment on the prevention and control of cucumber damping-off in potted plants

[0152] (1) Preparation of pathogens: Take out the PDA plate of Rhizoctonia solani stored at 4°C, use a 5-mm diameter puncher in a clean bench to punch out a round bacterial cake, transfer it to the center of the PDA medium plate, and incubate the inoculated PDA plate upside down in a 28°C incubator for 7 days for use.

[0153] (2) Preparation of fermentation broth: The fermentation broth was prepared according to step (1) of Experimental Example 5.

[0154] (3) Sowing: Select the cucumber seeds that germinated and grew evenly in Experimental Example 5 and transfer them to potted plants (11 cm in diameter, 9 cm in height, 7.5 cm in bottom diameter) with nutrient soil (containing a certain proportion of peat soil, vermiculite, perlite, etc.) sterilized at 121°C for 1 hour with high pressure steam. Inoculate one cucumber seed in each pot, and then cover the surface with 1-2 cm of sterilized nutrient soil. When the cucumber seedlings grow two true leaves, irrigate 30 mL of LangSG106 fermentation liquid near the roots of the cucumber seedlings. Use an equal volume of sterile water as a blank control group, and use an equal volume of 98% 2000-fold diluted solution of hydroxychloroquine soluble powder as a positive control group. Each treatment group has 12 pots, and each group has 12 cucumber seedlings.

[0155] (4) Pathogen inoculation: 24 hours after the FangSG106 fermentation liquid treatment group and the control group were inoculated, the roots of each seedling were pierced with a sterile needle and inoculated with 5 mm diameter Rhizoctonia solani fungi, 2 fungi per pot. Then, the plants were placed in a greenhouse with a temperature of 25±1°C, a relative humidity of 75%±5%, and a photoperiod of 14 hours of light / 10 hours of darkness to cultivate the disease. After 21 days, the soil at the roots of the cucumber seedlings was cleaned, and the incidence of the plants in the different treatment groups was counted to evaluate the severity of the disease. The disease index and relative prevention efficacy were calculated using the following formula.

[0156] Incidence rate = number of diseased plants / total number of plants × 100%

[0157] Disease index = 100% × ∑ (number of diseased plants at each level × representative value of each level) / (total number of plants surveyed × 4)

[0158] Relative control effect = (control disease index - treatment disease index) / control disease index × 100%

[0159] Table 5. Grading standards for cucumber damping-off disease

[0160]

[0161] Table 6. Effect of FangSG106 fermentation liquid on the prevention and control of cucumber damping-off disease

[0162]

[0163] The potted control effect is shown in Table 6. The results show that compared with the sterile water control group, the disease index of the two treatment groups decreased significantly. The disease index of the FangSG106 fermentation liquid treatment group was 16.72%, and the control efficiency was as high as 81.12%; while the disease index of the 98% oxazolidinone treatment group was 21.33%, and the control efficiency was 75.92%. The FangSG106 strain was slightly better than 98% oxazolidinone in controlling cucumber damping-off caused by Rhizoctonia solani.

[0164] Experimental Example 8: Potted Plant Control Experiment on Cucumber Wilt

[0165] (1) Preparation of pathogenic bacteria: Fusarium oxysporum was cultured on PDA at 28°C for 5 days, and then 10 bacterial blocks with a diameter of 5 mm were taken with a cork punch and transferred to a 500 mL Erlenmeyer flask containing 100 mL of PDB medium. The culture was incubated at 28°C in a shaker at 180 r / min for 3 days. The Fusarium oxysporum culture was filtered through 4 layers of sterile gauze, and the concentration of the filtered spore suspension was adjusted to 10 using a hemocytometer. 6 spores / mL.

[0166] (2) Preparation of fermentation broth: Fermentation broth was prepared according to step (1) of Experimental Example 5.

[0167] (3) Sowing: The sowing and treatment methods of cucumber seedlings are the same as those in step (3) of Experimental Example 7.

[0168] (4) Pathogen inoculation: 24 h after inoculation of FangSG106 fermentation broth in the treatment group and the control group, the root base of each seedling was pierced with a sterile needle and inoculated with 30 mL of 1×10 6 Spores / mL of Fusarium oxysporum spore suspension were then placed in a greenhouse with a temperature of 25±1°C, a relative humidity of 75%±5%, and a photoperiod of 14 hours of light / 10 hours of darkness to cultivate the disease. After 21 days, the soil at the roots of the cucumber seedlings was cleaned, and the incidence of the plants in different treatment groups was counted to evaluate the severity of the disease. The disease index and relative prevention effect were calculated using the following formula.

[0169] Incidence rate = number of diseased plants / total number of plants × 100%

[0170] Disease index = 100% × ∑ (number of diseased plants at each level × representative value of each level) / (total number of plants surveyed × 4)

[0171] Relative control effect = (control disease index - treatment disease index) / control disease index × 100%

[0172] Table 7. Grading standards for cucumber wilt disease

[0173]

[0174] Table 8. Effect of FangSG106 fermentation liquid on the control of cucumber wilt

[0175]

[0176] The potted control effect is shown in Table 8. The results show that compared with the sterile water control group, the disease index of the two treatment groups decreased significantly. The disease index of the FangSG106 fermentation liquid treatment group was 11.67%, and the control efficiency was as high as 85.71%; while the disease index of the 98% oxadiazon treatment group was 15.33%, and the control efficiency was 81.22%. The FangSG106 strain was slightly better than 98% oxadiazon in controlling cucumber wilt caused by Fusarium oxysporum.

[0177] Experimental Example 9: Determination of defensive enzyme activity in cucumber leaves

[0178] The leaves of the plants cultured for 21 days in Experimental Example 8 were collected and stored at -80°C. The activities of different defense enzymes in the plant leaves were determined using an enzyme activity assay kit. The five enzyme activities of catalase (CAT), polyphenol oxidase (PPO), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL) and peroxidase (POD) were determined using the detection kit purchased from Beijing Solebow Technology Co., Ltd. The detection method was referred to the kit instructions.

[0179] Table 9. Activities of five defense enzymes in cucumber leaves treated with FangSG106 fermentation liquid

[0180]

[0181] Note: The data in the table are mean ± standard error. Lowercase letters in the same column indicate significant differences in defense enzyme activities between the control group and the treatment group (P<0.05).

[0182] The experimental results are shown in Table 9. After 21 days of inoculation, the five enzyme activities of cucumber seedlings in the 98% oxadiazine positive control group and FangSG106 strain treatment group were significantly higher than those in the sterile water negative control group. Compared with the sterile water control group, the five enzyme activities of CAT, PPO, SOD, PAL and POD of cucumber seedlings inoculated with FangSG106 strain treatment increased by 116.27%, 93.40%, 94.18%, 84.30% and 108.77%, respectively.

[0183] Experimental Example 10: Antibacterial ability of Paenibacillus polymyxa FangSG106 against water pathogens

[0184] The Oxford cup method was used to detect the ability of Paenibacillus polymyxa FangSG106 to inhibit five aquatic pathogenic bacteria. The antibacterial experimental method is as follows:

[0185] (1) Preparation of aquatic pathogenic bacteria suspension:

[0186] Aeromonas vernix, Aeromonas spp. and Aeromonas punctatus were inoculated into TSB liquid medium at 2% inoculation volume, respectively, and cultured at 37°C and 220 rpm in a shaking incubator for 24 h. The bacterial solution concentration was adjusted to 10 8 CFU / mL is reserved.

[0187] Vibrio splendens and Vibrio anguillarum were inoculated into 2216E liquid culture medium at 2% inoculation volume, and cultured in a shaking incubator at 25°C and 220 rpm for 24 h. The bacterial solution concentration was adjusted to 10 8 CFU / mL is reserved.

[0188] (2) Antibacterial ability test:

[0189] The corresponding solid culture medium for pathogens (TSA solid culture medium for Aeromonas virginiana, Aeromonas scutellariae and Aeromonas punctata, 2216E solid culture medium for Vibrio splendens and Vibrio anguillarum) was cooled to about 50°C, and the pathogens were mixed into the culture medium in a certain proportion so that the number of viable bacteria of the pathogens was 10 6 CFU / mL order of magnitude, and then quickly poured into a plate with an Oxford cup placed in advance. After the culture medium cooled and solidified, the Oxford cup was taken out, 150 μL of Paenibacillus polymyxa FangSG106 fermentation liquid was added to the well, and the plates were placed in a constant temperature incubator at the corresponding temperature for 24-48 hours, and then the diameter of the inhibition zone was measured.

[0190] The antibacterial results are shown in Table 10. Paenibacillus polymyxa FangSG106 has a certain antibacterial ability against the five pathogenic bacteria.

[0191] Table 10. Antibacterial results of Paenibacillus polymyxa FangSG106 against five pathogenic bacteria

[0192]

Claims

1. A Paenibacillus polymyxa FangSG106, characterized in that: Its deposit number is CGMCC No.31655.

2. The use of Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655 in inhibiting bacteria, preventing and controlling plant diseases and promoting plant growth in non-therapeutic destinations.

3. The use of Paenibacillus polymyxa FangSG106 with the deposit number CGMCC No.31655 according to claim 2 in inhibiting bacteria in non-therapeutic destinations, preventing and controlling plant diseases, and promoting plant growth, characterized in that: The antibacterial spectrum includes: Rhizoctonia solani, Magnaporthe oryzae, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium oxysporum cucumber transformation, Fusarium wilt, Botrytis cinerea, Aeromonas verticillatus, Aeromonas jantaensis, Aeromonas punctatus, Vibrio splenici, and Vibrio anguillarum; And / or, the prevention of plant diseases refers to: preventing and controlling cucumber wilt and cucumber damping-off; And / or, the promoting plant growth refers to: promoting the growth of root length, plant height, stem thickness, fresh weight and dry weight of cucumber seedlings.

4. A biological pesticide comprising a pharmaceutically active ingredient, characterized in that: The pharmaceutically active ingredients include: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655.

5. A biological pesticide according to claim 4, characterized in that: Also includes: Pharmaceutically acceptable excipients.

6. A plant growth promoter comprising an active ingredient, characterized in that: The active ingredients include: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655.

7. A plant growth promoter according to claim 6, characterized in that: Also includes: Auxiliary materials.

8. A method for preventing and controlling plant diseases, characterized in that: Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655 is used to control plant diseases.

9. A method for promoting plant growth, characterized in that: Paenibacillus polymyxa FangSG106 with a deposit number of CGMCC No.31655 was used to promote plant growth.

10. A bacteriostatic method, characterized in that: Paenibacillus polymyxa FangSG106 with a preservation number of CGMCC No.31655 was used for antibacterial treatment.

Images