Bacillus velezensis MSGL-ZJ1 as well as application and microbial preparation thereof

Abstract

The invention relates to the technical field of microbial engineering, and particularly discloses bacillus velezensis MSGL-ZJ1 as well as application and a microbial preparation thereof. The bacillus velezensis MSGL-ZJ1 is preserved in the China General Microbiological Culture Collection Center on September 30, 2024, and the preservation number of the bacillus velezensis MSGL-ZJ1 is CGMCC (China General Microbiological Culture Collection Center) NO.32125. The bacillus velezensis MSGL-ZJ1 is named as bacillus velezensis MSGL-ZJ1. The bacillus velezensis MSGL-ZJ1 and the fermentation liquor thereof are applied to preparation of products for inhibiting microorganisms or / and promoting plant growth. The microbial composition is prepared from the bacillus velezensis MSGL-ZJ1, and further comprises the bacillus velezensis MSGL-ZJ1. The bacillus velezensis MSGL-ZJ1 and the microbial composition provided by the invention can effectively inhibit the growth of phytopathogens, can be used as a broad-spectrum antibacterial biocontrol inoculant, and can effectively promote the growth and development of plants and enhance the stress resistance of the plants.

Description

Technical Field

[0001] The invention relates to the technical field of microbial engineering, in particular to Bacillus velez MSGL-ZJ1 and applications thereof and microbial preparations. Background Art

[0002] Bacillus belongs to the Bacillaceae family and the genus Bacillus. It is a type of aerobic or facultative anaerobic, endospore-forming Gram-positive rod. It is widely distributed in water, soil, air, plant roots, plant surfaces and animal intestines in nature. It has good characteristics of strong stress resistance, rapid growth and stable performance. Its products are rich in proteases, cellulases and other active substances, which help regulate the balance of intestinal flora, degrade toxic substances, improve feed conversion rate, and enhance the body's immunity. It is often used as a feed additive for animal production.

[0003] Biological control of plant diseases mainly refers to the technology of using beneficial microorganisms and their metabolites to prevent and control diseases. In essence, it is to use the direct or indirect competition, antibiotics and bacteriolysis of microorganisms, or to use hormones and other substances produced by microorganisms to resist pathogens, thereby achieving the effect of inhibiting pathogens. Summary of the invention

[0004] The purpose of the present invention is to solve the shortcomings in the prior art and to propose Bacillus Velez MSGL-ZJ1 and its application and microbial preparation.

[0005] Bacillus Velez Bacillus velezensis )MSGL-ZJ1, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on September 30, 2024, with the deposit number CGMCC NO.32125.

[0006] Preferably, the 16S rRNA gene sequence of Bacillus velez MSGL-ZJ1 is shown as SEQ ID NO.1.

[0007] The invention discloses a Bacillus Velez subsp. MSGL-ZJ1 fermentation liquid, which is obtained by fermenting the Bacillus Velez subsp. MSGL-ZJ1.

[0008] The preparation method of the fermentation broth of Bacillus Velez MSGL-ZJ1 comprises the following steps: inoculating Bacillus Velez MSGL-ZJ1 into NB medium, culturing at 25-28°C for 60-84h, subculturing 1-3 times, washing by centrifugation, and adjusting the system concentration to its OD 600nm to 0.8-1.1, and then inoculated into new NB medium and cultured at 25-28℃ for 60-84h.

[0009] The above-mentioned Bacillus Velez MSGL-ZJ1 is used in preparing products for inhibiting microorganisms or / and promoting plant growth.

[0010] The above-mentioned Bacillus Velez MSGL-ZJ1 fermentation liquid is used in preparing products for inhibiting microorganisms or / and promoting plant growth.

[0011] Preferably, the inhibited microorganism is at least one of Gardnerella vaginalis, Porphyromonas gingivalis, Candida albicans, Streptococcus mutans, Staphylococcus aureus, Sclerotium solani, Helminthosporium, and Fusarium moniliforme.

[0012] Preferably, the function of promoting plant growth is achieved by increasing the content of soluble phosphorus and / or indoleacetic acid in the system.

[0013] A microbial composition comprises: the above-mentioned Bacillus Velez subtilis MSGL-ZJ1.

[0014] Preferably, the above-mentioned microbial composition further comprises: Lactobacillus plantarum ( Lactiplantibacillus plantarum )MLL-6, Lactobacillus paracasei ( Lacticaseibacillus paracasei )RBB-10.

[0015] Preferably, in the above microbial composition, the concentration of live bacteria of Bacillus Velezii MSGL-ZJ1 is 4-8×10 9 cfu / g, and the live bacterial concentration of Lactobacillus plantarum MLL-6 was 7-9×10 8 cfu / g, and the live bacterial concentration of Lactobacillus paracasei RBB-10 was 3-7×10 8 cfu / g.

[0016] A microbial preparation is obtained by anaerobic fermentation of the above microbial composition.

[0017] Preferably, the anaerobic fermentation temperature is 28-37°C, and the anaerobic fermentation time is 72-144h. Beneficial effects: The present invention provides the above-mentioned Bacillus Velezii MSGL-ZJ1, which has obvious inhibitory effects on vaginal Gardnerella, Candida albicans (Gardnerella and Candida albicans are the main pathogens causing female vaginal diseases), Porphyromonas gingivalis, Streptococcus mutans (Porphyromonas gingivalis and Streptococcus mutans are the main pathogens causing dental health), Staphylococcus aureus (the most common pathogen in human suppurative infection), Sclerotium solani (the main pathogen causing white rot), Helminthosporium (the main pathogen causing corn leaf spot), and Fusarium moniliforme (the main pathogen causing seedling blight).

[0018] The diameters of the inhibition zones of the fermentation liquid of Bacillus Velez MSGL-ZJ1 against Gardnerella vaginalis, Porphyromonas gingivalis, Candida albicans, Streptococcus mutans, and Staphylococcus aureus were 1.80±0.02cm, 2.15±0.05cm, 1.50±0.03cm, 1.70±0.04cm, and 1.15±0.01cm, respectively; while the inhibition rates against Microsclerotium alignum, Helminthosporium longifolium, and Fusarium moniliforme were 86.42%, 85.33%, and 60.47%, respectively.

[0019] The applicant has found that the combination of Bacillus Velezii MSGL-ZJ1 with Lactobacillus plantarum MLL-6 and Lactobacillus paracasei RBB-10 exhibits good synergistic behavior, and the obtained microbial composition is subjected to anaerobic fermentation, so that the inhibitory effect of the obtained fermentation liquid on Gardnerella vaginalis, Porphyromonas gingivalis, Candida albicans, Streptococcus mutans, Staphylococcus aureus, Sclerotium uniformis, Helminthosporium longifolium and Fusarium moniliforme is further enhanced.

[0020] The above-mentioned Bacillus Velezii MSGL-ZJ1 and microbial composition provided in the present application can effectively inhibit the growth of plant pathogens, can be used as a broad-spectrum antibacterial biocontrol agent, and can effectively promote plant growth and development and enhance plant resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a photo of the colony morphology of Bacillus Velez MSGL-ZJ1.

[0022] Figure 2 This is a microscopic photo of Bacillus Velez MSGL-ZJ1.

[0023] Figure 3 These are photos of the antibacterial test of Bacillus Velez MSGL-ZJ1 fermentation broth against Gardnerella vaginalis.

[0024] Figure 4 These are photos of the antibacterial test of Bacillus Velez MSGL-ZJ1 fermentation broth against Porphyromonas gingivalis.

[0025] Figure 5 These are photos of the antibacterial test of Bacillus Velez MSGL-ZJ1 fermentation broth against Streptococcus mutans.

[0026] Figure 6 These are photos of the antibacterial test of Bacillus Velez MSGL-ZJ1 fermentation broth against Staphylococcus aureus.

[0027] Figure 7 Schematic diagram of the confrontation experiment between Bacillus velez MSGL-ZJ1 fermentation broth and Sclerotium sclerotiorum.

[0028] Figure 8These are photos of the antibacterial test of Bacillus Velez MSGL-ZJ1 fermentation broth against Sclerotium microsporum.

[0029] Fig. 9 These are photos of the antibacterial test of the fermentation broth of Bacillus Velez MSGL-ZJ1 against Helminthosporium.

[0030] Fig.10 These are photos of the antibacterial test of the fermentation broth of Bacillus Velez MSGL-ZJ1 against Fusarium moniliforme.

[0031] Fig.11 This is a photo of the starch decomposition experiment of Bacillus Velez MSGL-ZJ1.

[0032] Fig.12 This is a photo of the cellulose decomposition experiment of Bacillus Velez MSGL-ZJ1.

[0033] Fig.13 This is the effective phosphorus capacity curve of Bacillus Velez MSGL-ZJ1.

[0034] Fig.14 This is the indoleacetic acid production ability curve of Bacillus Velez MSGL-ZJ1.

[0035] Fig.15 This is a comparison chart of the inhibition zone diameters and inhibition zone diameter growth rates of the fermentation broth of Bacillus Velez MSGL-ZJ1 and the microbial composition against Gardnerella vaginalis, Candida albicans, Porphyromonas gingivalis, Streptococcus mutans, and Staphylococcus aureus.

[0036] Fig.16 It is a comparison chart of the inhibition rate and inhibition rate growth rate of the fermentation broth of Bacillus Velezii MSGL-ZJ1 and the microbial composition against Sclerotium microspore, Helminthosporium and Fusarium moniliforme. DETAILED DESCRIPTION

[0037] The present invention will be further explained below in conjunction with specific embodiments.

[0038] The following Lactobacillus plantarum ( Lactiplantibacillus plantarum ) The deposit number of MLL-6 is CGMCC NO.30469, which is isolated by the applicant (Hangzhou Junwu Xiyan Biotechnology Co., Ltd.).

[0039] The following Lactobacillus paracasei ( Lacticaseibacillus paracasei ) The deposit number of RBB-10 is CGMCC NO.30557, which is isolated by the applicant (Hangzhou Junwu Xiyan Biotechnology Co., Ltd.).

[0040] The NB culture medium used below was purchased from Qingdao Hibo Biotechnology with the brand name HB0108.

[0041] The NA medium used below was obtained by adding 2% agar to the above NB medium.

[0042] The TSA culture medium used below was purchased from Qingdao Hibo Biotechnology with the brand number HB0177-1.

[0043] The defibrinated sheep blood used in the following was purchased from Qingdao Hibo Biological Technology Co., Ltd., with the brand number 1001339-1.

[0044] The PDA culture medium used below was purchased from Qingdao Hibo Biotechnology Co., Ltd., with the brand number HB0233-12.

[0045] The inorganic phosphorus culture medium used below was purchased from Qingdao Hibo Biotechnology with the brand name HB8670-1.

[0046] The organophosphorus culture medium used below was purchased from Qingdao Hibo Biotechnology with the brand number HB8673-1.

[0047] Example 1: Separation, purification, and identification This strain was isolated and collected by Huang Dasheng from homemade fermentation broth in Shangcheng District, Hangzhou City, Zhejiang Province, China in June 2024.

[0048] Isolation and purification of strains: From the homemade fermentation broth sample, 100 μL of the sample was taken into 900 μL of sterile water, and then the sample was diluted to 10 -4 , 10 -5 , place in a 70℃ water bath for 20 min, spread 100 μL of the liquid after the water bath on a NA plate, and place in a 28℃ incubator for 72 h.

[0049] Observe the bacterial growth on the plate. After the colonies grow, pick a typical single colony and streak it on the NA solid medium for strain isolation and purification. Inoculate the single colony into 5 mL of sterilized NB medium, incubate at 28°C for 72 hours, and then store it in a -80°C refrigerator with 50% glycerol for later use.

[0050] The isolated strains were identified by morphology (colony morphology such as Figure 1 As shown, the microscopic morphology Figure 2 as shown).

[0051] The isolated and purified single colonies were sent to Zhejiang Shangya Biotechnology Co., Ltd. for sequencing identification. The primers are as follows: 27F: 5'-AGAGTTTGATCMTGGCTCAG-3'; 1552R: 5'-AAGGAGGTGATCCAGCCGCA-3'.

[0052] Its 16s rDNA sequence: The BLAST program in NCBI was used to compare the sequences. The results showed that the MSGL-ZJI strain was Bacillus Velezii, and its homology with the known Bacillus in the GeneBank database reached 99.93%. It was named Bacillus Velezii ( Bacillus velezensis )MSGL-ZJ1.

[0053] Bacillus velez MSGL-ZJ1 was deposited in the General Microbiology Center of China Culture Collection Administration (Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) on ​​September 30, 2024, with the deposit number CGMCC NO.32125.

[0054] Example 2: Antibacterial effect on Gardnerella vaginalis 2.1 Prepare the Tryptic Soy Blood Agar plates (made with TSA medium and defibrinated sheep blood) in advance and let them dry.

[0055] 2.2 Streak the activated vaginal Gardnerella (SHBCC) on the above tryptic soy blood agar plate ® , ATCC49145), then pick a single colony into a modified TSB medium (i.e., TSB with 5% serum), culture at 37°C for 5-10 days, then transfer to a new modified TSB medium at a ratio of 2%, and continue to culture at 37°C for 5-10 days to obtain a fresh bacterial solution. Subsequently, centrifuge the bacterial solution at 5000r / min for 5 minutes, remove the supernatant, add PBS buffer to wash the bacteria twice, and the resulting bacterial solution is used for later use.

[0056] 2.3 The Bacillus Velez MSGL-ZJ1 bacterial suspension in the cryopreserved tube was inoculated into NB medium at a 2% inoculation rate and cultured in a 28°C incubator for 72 h. After subculturing twice, the bacteria were washed twice by centrifugation (5000 r / min for 5 min) with PBS buffer, and then the OD value of the strain was calculated by using PBS buffer. 600nm Adjust to 1.0. Inoculate the strain into NB medium at 2% inoculation amount, culture at 28℃ for 72h, and the obtained fermentation broth is used for later use.

[0057] 2.4 Pour a thin layer of NA medium at the bottom of the culture dish and put in 3 Oxford cups. Add vaginal Gardnerella to the modified TSA medium at a volume ratio of 10% in advance, mix well, then pour the medium mixed with vaginal Gardnerella into the culture dish. After the medium solidifies, take out the Oxford cup. Add 100 μL of the fermentation liquid of the above-mentioned Bacillus Velezii MSGL-ZJ1 strain into each well, culture anaerobically at 28℃ for 5 days, and observe the inhibition zone.

[0058] The results show (such as Figure 3As shown in the figure, Bacillus Velezii MSGL-ZJ1 has a significant inhibitory effect on vaginal Gardnerella, and the diameter of the inhibition zone produced by its fermentation broth is 1.80±0.02cm. However, when streptomycin (antibiotic) with a concentration of 20μg / mL was used as a control, no inhibition zone was produced, and it had no inhibitory effect on vaginal Gardnerella.

[0059] Example 3: Antibacterial effect on Porphyromonas gingivalis 3.1 Streaking of Porphyromonas gingivalis (SHBCC) on tryptic soy blood agar plates ® , ATCC 33277), then pick a single colony into a modified TSB medium (i.e., TSB with 5% serum), culture at 37°C anaerobically for 5-10 days, then transfer to a new modified TSB medium at a ratio of 2%, culture at 37°C anaerobically for 5-10 days, and obtain a fresh bacterial solution. Subsequently, centrifuge the bacterial solution at 5000r / min for 5min to remove the supernatant, add PBS buffer to wash the bacteria twice, and the resulting bacterial solution is used for later use.

[0060] 3.2 The Bacillus Velez MSGL-ZJ1 bacterial suspension in the cryopreserved tube was inoculated into NB medium at a 2% inoculation rate and cultured in a 28°C incubator for 72 h. After subculturing twice, the bacteria were washed twice by centrifugation (5000 r / min for 5 min) with PBS buffer, and then the strain OD was increased by 1.5% using PBS buffer. 600nm Adjust to 1.0, inoculate the strain into NB medium at a 2% inoculation rate, culture at 28°C for 3 days, and the resulting fermentation broth is ready for use.

[0061] 3.3 Pour a thin layer of NA medium at the bottom of the culture dish and put in 3 Oxford cups. Add Porphyromonas gingivalis to the modified TSA medium at a volume ratio of 10% in advance, mix well, then pour the medium mixed with Porphyromonas gingivalis into the culture dish. After the medium solidifies, take out the Oxford cup. Add 100 μL of the fermentation liquid of the above-mentioned Bacillus Velezii MSGL-ZJ1 strain into each well, culture anaerobically at 28℃ for 5 days, and observe the inhibition zone.

[0062] The results show (such as Figure 4 As shown in the figure, Bacillus Velezii MSGL-ZJ1 has a significant inhibitory effect on Porphyromonas gingivalis, and the diameter of the inhibition zone produced by its fermentation broth is 2.15±0.05cm. However, when streptomycin (antibiotic) with a concentration of 20μg / mL was used as a control, no inhibition zone was produced, and it had no inhibitory effect on Porphyromonas gingivalis.

[0063] Example 4: Antibacterial effect on Candida albicans, Streptococcus mutans, and Staphylococcus aureus Referring to the Oxford cup method in Example 2 and Example 3, antibacterial experiments were carried out on Candida albicans, Streptococcus mutans and Staphylococcus aureus.

[0064] The results show (such as Figure 5 and Figure 6 As shown in the figure, Bacillus Velezii MSGL-ZJ1 has an inhibitory effect on Candida albicans, Streptococcus mutans, and Staphylococcus aureus, and the diameters of the inhibition zones produced by its fermentation broth are 1.50±0.03cm, 1.70±0.04cm, and 1.15±0.01cm, respectively. However, when streptomycin (antibiotic) with a concentration of 20μg / mL was used as a control, no inhibition zone was produced, and it had no inhibitory effect on the above-mentioned bacteria.

[0065] Gardnerella and Candida albicans are the main pathogens that cause female vaginal diseases, Porphyromonas gingivalis and Streptococcus mutans are the main pathogens that cause dental health, and Staphylococcus aureus is the most common pathogen in human suppurative infections, which can cause local suppurative infections. Bacillus Velezii MSGL-ZJ1 has a significant inhibitory effect on the above-mentioned human pathogens, pathogens that cause female vaginal diseases and dental diseases.

[0066] Example 5: Antibacterial effect on white rot 5.1 Prepare potato dextrose agar plates (PDA medium) in advance and let them dry.

[0067] 5.2 The Bacillus Velez MSGL-ZJ1 bacterial suspension in the cryopreserved tube was inoculated into NB medium at a 2% inoculation rate and cultured in a 28°C incubator for 72 h. After subculturing twice, the bacteria were washed twice by centrifugation (5000 r / min for 5 min) with PBS buffer, and then the strain OD was increased by 1.5% using PBS buffer. 600nm Adjust to 1.0, inoculate the strain into NB medium at a 2% inoculation rate, culture at 28°C for 72h, and the resulting fermentation broth is ready for use.

[0068] 5.3 If Figure 7 As shown, 4 Oxford cups (i.e. Figure 7 Then pour the PDA medium into the culture dish, wait for the medium to solidify, and take out the Oxford cup. Add 100 μL of the fermentation liquid of the above-mentioned Bacillus Velez MSGL-ZJ1 strain into each well.

[0069] Finally, place a neat block of small sclerotium of the same size in the center of the solidified plate (which is the main pathogen causing white rot). For the blank control, only a neat block of small sclerotium (i.e. Figure 7 The culture was carried out at 28°C for about 5 days, and the growth of Sclerotium was observed.

[0070] Inhibition rate = (natural growth diameter of Sclerotium uniformis - growth diameter of Sclerotium uniformis after being inhibited by Bacillus Velezii MSGL-ZJ1 fermentation broth) ÷ natural growth diameter of Sclerotium uniformis × 100% The results show (such as Figure 8 As shown in the figure), Bacillus Velezii MSGL-ZJ1 had a significant inhibitory effect on Sclerotium oxysporum, and the inhibition rate of its fermentation broth on Sclerotium oxysporum reached 86.42%.

[0071] Example 6: Antibacterial effect on corn leaf spot and seedling blight Similarly, referring to the confrontation experimental method in Example 5, antibacterial experiments were conducted on Helminthosporium (which is the main pathogen causing corn leaf blight) and Fusarium moniliforme (which is the main pathogen causing seedling blight).

[0072] The results show (such as Fig. 9 and Fig.10 As shown in the figure), Bacillus Velezii MSGL-ZJ1 had a significant inhibitory effect on Helminthosporium and Fusarium moniliforme, and the inhibition rates of its fermentation broth on Helminthosporium and Fusarium moniliforme were 85.33% and 60.47%, respectively.

[0073] Example 7: Determination of the ability to decompose starch and cellulose 7.1 Use the spot method to inoculate Bacillus Velez MSGL-ZJ1 on the starch screening medium (3 replicates) and culture it in a 28℃ incubator for 5 days. Then add dilute iodine solution to the medium to observe whether there is a hydrolysis circle.

[0074] The composition of starch screening medium is as follows: soluble starch 10g, peptone 2g, yeast extract 5g, NaCl 5g, K2HPO4 3g, agar 20g, add water to make up to 1000mL.

[0075] like Fig.11 As shown, Bacillus Velez MSGL-ZJ1 has the ability to decompose starch.

[0076] 7.2 Inoculate Bacillus velezensis MSGL-ZJ1 on cellulose screening medium (3 replicates) by spot seeding method and culture at 28℃ for 5 days. Pour 1mg / mL Congo red solution into the culture dish for staining for 30 minutes, pour out the Congo red solution, soak in 1mol / L NaCl solution for 15 minutes for decolorization, observe whether there is a hydrolysis circle and measure the diameter of the hydrolysis circle.

[0077] The composition of the cellulose screening medium is as follows: sodium carboxymethyl cellulose 10.0 g, (NH4)2SO4 1.0 g, MgSO4•7H2O0.5 g, KH2PO4 1.0 g, NaCl 0.3 g, agar powder 20.0 g, add water to make up to 1000 mL.

[0078] like Fig.12 As shown, Bacillus Velez MSGL-ZJ1 has the ability to decompose cellulose, and the diameter of its hydrolysis zone is 2.7±0.15 cm.

[0079] Bacillus can degrade polysaccharides in fungal cell walls by secreting enzymes (such as cellulase and amylase), destroy the structure of pathogens, degrade fungal hyphae, inhibit the growth of pathogens, and ultimately lead to the death of pathogens.

[0080] Example 8: Determination of Available Phosphorus Capacity The OD of the suspension of Bacillus velezensis MSGL-ZJ1 was 600nm Adjust to 1.0, and then inoculate into 50mL centrifuge tubes containing 30mL of organic phosphorus liquid culture medium and inorganic phosphorus liquid culture medium at an inoculum size of 2%, respectively. The organic phosphorus liquid culture medium and inorganic phosphorus liquid culture medium without inoculation of strains were used as blank controls (CK).

[0081] The cells were cultured at 28°C with shaking at 200 rpm for 5 days, and samples were taken at corresponding time points (24 h, 48 h, 72 h, 96 h, and 120 h). 5 mL of the sample was collected each time and centrifuged at 4°C at 10,000 r / min for 5 min. The supernatant was stored at 4°C (repeated 3 times).

[0082] The molybdenum antimony colorimetric method was used to determine the soluble phosphorus content in the supernatant at 24h, 48h, 72h, 96h and 120h.

[0083] like Fig.13 As shown, Bacillus Velez MSGL-ZJ1 effectively decomposes inorganic phosphorus and promotes phosphorus to exist in the form of soluble phosphorus, which is convenient for plant absorption and promotes plant growth.

[0084] Example 9: Determination of the ability to produce indoleacetic acid (IAA) The OD of the suspension of Bacillus velezensis MSGL-ZJ1 was 600nm Adjust to 1.0, then inoculate 2% of the inoculum into a 10mL test tube containing 5mL NB liquid medium (containing 100mg / L L-tryptophan), and place in a shaker at 28℃ 200rpm for 5 days (3 replicates). Take 500μL of the bacterial solution cultured at different times (24h, 48h, 72h, 96h, 120h) into a 1.5mL EP tube, centrifuge at 12000rpm for 5min, and keep the supernatant for later use.

[0085] Take 50 μL of supernatant in a 96-well plate and add 50 μL of Salkowski colorimetric solution (mixture of solution A and solution B). After the 96-well plate is placed in the dark at room temperature for 30 minutes, the OD is measured using an enzyme-labeled instrument. 530nm The IAA concentration was calculated from the absorbance value.

[0086] like Fig.14 As shown, Bacillus Velezii MSGL-ZJ1 produces indoleacetic acid. As time goes by, the concentration of indoleacetic acid shows a slow upward trend as a whole, which can promote the growth and development of the plant.

[0087] Example 10: Microbial composition fermentation broth The bacterial solution of Bacillus Velez MSGL-ZJ1, the bacterial solution of Lactobacillus plantarum MLL-6, and the bacterial solution of Lactobacillus paracasei RBB-10 were mixed, and the live bacterial concentration of Bacillus Velez MSGL-ZJ1 in the system was 6×10 9 cfu / g, and the live bacterial concentration of Lactobacillus plantarum MLL-6 was 8×10 8 cfu / g, and the live bacterial concentration of Lactobacillus paracasei RBB-10 was 4×10 8 cfu / g.

[0088] The mixed bacterial liquid was inoculated into a 5L small anaerobic fermentation tank at a 2% inoculation rate. The temperature in the fermentation tank was adjusted to 37±1°C. The fermentation was cultured for 72 hours. Fresh culture medium was added. Then the temperature in the fermentation tank was adjusted to 28±1°C. The fermentation was cultured for 72 hours. The obtained fermentation liquid was used for later use.

[0089] Example 11: Antibacterial effect of fermented liquid of microbial composition on Gardnerella vaginalis, Candida albicans, Porphyromonas gingivalis, Streptococcus mutans, and Staphylococcus aureus The antibacterial experiments were carried out on Gardnerella vaginalis, Candida albicans, Porphyromonas gingivalis, Streptococcus mutans and Staphylococcus aureus by using the Oxford cup method in Reference Example 2-4, the diameter of the antibacterial zone was measured, and the growth rate of the diameter of the antibacterial zone of the fermentation liquid of the microbial composition was calculated.

[0090] Growth rate of inhibition zone diameter = (diameter of inhibition zone of microbial composition fermentation liquid - diameter of inhibition zone of Bacillus Velez MSGL-ZJ1 fermentation liquid) ÷ diameter of inhibition zone of Bacillus Velez MSGL-ZJ1 fermentation liquid × 100% like Fig.15As shown, the microbial composition has an inhibitory effect on Gardnerella vaginalis, Candida albicans, Porphyromonas gingivalis, Streptococcus mutans, and Staphylococcus aureus. Compared with Bacillus Velezii MSGL-ZJ1, the growth rate of the diameter of the inhibition zone of the fermentation liquid of the microbial composition is at least 50%, and the highest can reach 73.33%.

[0091] Example 12: Antibacterial effect of the fermentation broth of the microbial composition on Sclerotium sclerotiorum, Helminthosporium and Fusarium moniliforme Referring to the confrontation experimental method in Example 5, antibacterial experiments were carried out on Sclerotium sclerotiorum, Helminthosporium, and Fusarium moniliforme.

[0092] Antibacterial rate growth rate = (antibacterial rate of microbial composition fermentation liquid - antibacterial rate of Siamese Bacillus PHBM2-Ba5 fermentation liquid) ÷ antibacterial rate of Siamese Bacillus PHBM2-Ba5 fermentation liquid × 100% like Fig.16 As shown, the microbial composition has an inhibitory effect on Sclerotium uniformis, Helminthosporium and Fusarium moniliforme. Compared with Bacillus velezensis MSGL-ZJ1, the growth rate of the microbial composition fermentation liquid is at least 10%, and the highest can reach 20%.

[0093] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. Bacillus Velez Bacillus velezensis )MSGL-ZJ1, characterized in that, It was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on September 30, 2024, with the deposit number CGMCC NO.32125.

2. The Bacillus Velezii MSGL-ZJ1 according to claim 1, characterized in that Its 16S rRNA gene sequence is shown in SEQ ID NO.

1.

3. A fermentation broth of Bacillus Velez MSGL-ZJ1, characterized in that: The product is obtained by fermenting the Bacillus Velez MSGL-ZJ1 described in claim 1 or 2.

4. A method for preparing the fermentation broth of Bacillus Velez MSGL-ZJ1 according to claim 3, characterized in that: The method comprises the following steps: inoculating Bacillus velez MSGL-ZJ1 in NB medium, culturing at 25-28°C for 60-84h, subculturing 1-3 times, washing by centrifugation, and adjusting the concentration of the system to its OD 600nm to 0.8-1.1, and then inoculated into new NB medium and cultured at 25-28℃ for 60-84h.

5. Use of the Bacillus Velez MSGL-ZJ1 according to claim 1 or the fermentation liquid of the Bacillus Velez MSGL-ZJ1 according to claim 3 in preparing products for inhibiting microorganisms and / or promoting plant growth.

6. The use according to claim 5, characterized in that: The microorganism is at least one of Gardnerella vaginalis, Porphyromonas gingivalis, Candida albicans, Streptococcus mutans, Staphylococcus aureus, Sclerotium uniformis, Helminthosporium longisporum, and Fusarium moniliforme.

7. The use according to claim 5, characterized in that: The function of promoting plant growth is achieved by increasing the content of soluble phosphorus and / or indoleacetic acid in the system.

8. A microbial composition, characterized in that include: The Bacillus Velez MSGL-ZJ1 as claimed in claim 1.

9. The microbial composition according to claim 8, characterized in that: Also includes: Lactobacillus plantarum ( Lactiplantibacillus plantarum )MLL-6, Lactobacillus paracasei ( Lacticaseibacillus paracasei )RBB-10; The concentration of live bacteria of Bacillus velez MSGL-ZJ1 is 4-8×10 9 cfu / g, and the live bacterial concentration of Lactobacillus plantarum MLL-6 was 7-9×10 8 cfu / g, and the live bacterial concentration of Lactobacillus paracasei RBB-10 was 3-7×10 8 cfu / g.

10. A microbial preparation, characterized in that: Obtained by anaerobic fermentation of the microbial composition according to claim 8 or 9.

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