Bacillus amyloliquefaciens with tomato disease control function and application of bacillus amyloliquefaciens
By using Bacillus amyloliquefaciens ZN-S18 and its metabolites, the problem of chemical contamination in tomato disease control has been solved, achieving effective inhibition of various pathogens, enhancing the disease resistance of tomato plants, and demonstrating broad application prospects.
Patent Information
- Application Number
- CN202511599340.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-04
- Publication Date
- 2026-02-06
AI Technical Summary
In existing technologies, the control of tomato diseases mainly relies on chemical agents, which poses risks such as soil pollution, pathogen resistance, and ecological harm. Furthermore, the ability of isolated strains to colonize within plants is insufficient, limiting their practical application effectiveness.
The Bacillus amyloliquefaciens strain ZN-S18 was used to prepare metabolites such as surfactant and subtilisin to inhibit the growth and spore germination of Fusarium oxysporum, Botrytis cinerea, and Ralstonia solanacearum, and was applied to control Fusarium wilt and bacterial wilt in tomato seedlings.
The ZN-S18 strain exhibits significant broad-spectrum antagonistic activity in the control of tomato diseases, effectively inhibiting a variety of pathogens, improving the disease resistance of tomato plants, and demonstrating good potential for biological control and environmental friendliness.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of crop disease prevention and control, in particular to a bacillus amyloliquefaciens with tomato disease prevention and treatment function and application thereof. BACKGROUND
[0002] Tomato (Solanum lycopersicum) is one of the most important economic crops in the world, and plays an irreplaceable role in agricultural production and food industry. According to the statistics of the Food and Agriculture Organization of the United Nations (FAO, 2023), the global annual output of tomatoes is as high as 192 million tons, with China ranking first in the world with an annual output of 70 million tons, accounting for 30% of the global total. The planting area and output have maintained steady growth. In 2021, the planting area of tomatoes in China reached 1,111,500 hectares, with an output value of over 180 billion yuan. Tomatoes not only have important economic value, but also are rich in vitamins C and lycopene and other antioxidants, and can be eaten fresh or processed, becoming an indispensable part of the residents' dietary structure.
[0003] However, with the expansion of tomato planting scale, diversification of cultivars and increasing complexity of cultivation patterns, disease problems have become increasingly serious. According to literature reports, tomatoes are susceptible to more than 200 diseases, of which soil-borne diseases are particularly prominent. Tomato wilt caused by Fusarium oxysporum f. sp. lycopersici is one of the main diseases in facilities and open field cultivation, which can cause severe yield reduction, with a loss rate of more than 80% in some areas. The pathogen invades and expands to the vascular bundle through the root epidermis, blocks water transport in the conduit, and then causes leaf yellowing, vascular browning and plant wilting, eventually leading to whole plant death. More troublesome is that the pathogen can form dormant chlamydospores under adverse conditions, survive in the soil for a long time and occupy a dominant position, making it a persistent pathogen difficult to eradicate.
[0004] In addition to fungal diseases, tomato bacterial wilt caused by Ralstonia solanacearum is also devastating. The optimal temperature for the pathogen to cause disease is 20-38℃, and it is often found in high-temperature and high-humidity environments. In recent years, affected by global warming and changes in planting patterns, the disease has shown an increasing trend in occurrence in temperate and cool regions of China, especially in South China and the Yangtze River Basin, with a disease incidence of up to 80%, seriously restricting the development of the tomato industry. Ralstonia solanacearum invades the root and rapidly expands to the vascular bundle, causing acute wilting of the plant, with symptoms particularly evident in the stem vascular bundle, ultimately causing large-scale yield reduction. In addition, air-borne pathogens such as Botrytis cinerea are also prevalent in tomato production, posing a serious threat to yield and quality.
[0005] At present, chemical agents are still the main means of tomato disease prevention and control, including irrigation, spraying and soil treatment and other measures. Although chemical control can quickly take effect in the short term, its disadvantages are increasingly prominent: long-term use not only leads to soil pollution and water eutrophication, but also promotes the development of drug resistance of pathogenic bacteria, and causes potential harm to non-target organisms and the ecological environment. These problems have become a prominent bottleneck restricting the sustainable development of agriculture. Therefore, the development of safe, efficient and environmentally friendly alternative control measures has become an inevitable trend of tomato disease integrated management.
[0006] Among various new control strategies, biological control has attracted widespread attention due to its advantages such as not easy to produce resistance, long effective period, and small environmental pollution. Among them, plant endophyte is considered as a potential biological control resource library. Endophyte can often stably colonize in plant tissue during long-term symbiosis with the host, and inhibit pathogenic bacteria through various ways such as producing antibacterial substances, competing for nutrients and ecological niche, and inducing host systemic resistance, which has good application prospect. However, the isolation of most antagonistic bacteria is concentrated in the rhizosphere and roots of plants, and the microbial community in these areas is extremely complex. Although some isolated strains show strong antagonistic activity, whether they are true endophytes is still doubtful, and their colonization ability in plant body is insufficient, thereby limiting their application effect in actual production.
[0007] Therefore, a bacillus amyloliquefaciens with tomato disease control function and its application are provided. SUMMARY
[0008] TECHNICAL PROBLEMS SOLVED
[0009] In view of the deficiencies in the prior art, the present application provides a bacillus amyloliquefaciens with tomato disease control function and its application. The ZN-S18 strain has a significant effect in the prevention and control of tomato wilt, and also shows broad-spectrum antagonistic activity in inhibiting various pathogenic bacteria, including the mycelial growth of fusarium oxysporum (tomato special type and strawberry special type), colletotrichum gloeosporioides, botrytis cinerea, and the proliferation of pseudomonas solanacearum. In addition, ZN-S18 can also significantly inhibit the spore germination of fusarium oxysporum (tomato special type) and the sporulation of fusarium oxysporum and botrytis cinerea. The present application solves the problem that the colonization ability of isolated strains in plant body is insufficient, thereby limiting their application effect in actual production.
[0010] TECHNICAL SCHEME
[0011] To achieve the above-mentioned purpose, the present application provides the following technical scheme:
[0012] The application discloses a bacillus amyloliquefaciens with a function of preventing and treating tomato diseases, which is identified as bacillus amyloliquefaciens ZN-S18, and has been preserved in the China General Microbiological Culture Collection Center on September 9, 2025, with a preservation number of CGMCC No.35860 and an address of No.3, Institute of Microbiology, Chinese Academy of Sciences, Beijing.
[0013] A biocontrol agent:
[0014] The biocontrol agent comprises bacillus amyloliquefaciens ZN-S18.
[0015] Preferably, the preparation method of the biocontrol agent comprises the following steps:
[0016] Step one: inoculating activated bacillus amyloliquefaciens ZN-S18 into CM culture solution and culturing for 24 hours;
[0017] Step two: when the OD value of the culture solution reaches 1.0-1.5, the biocontrol agent is obtained. 600
[0018] A metabolite:
[0019] The metabolite is a metabolite of bacillus amyloliquefaciens ZN-S18.
[0020] Preferably, the ZN-S18 metabolite comprises two types of cyclic lipopeptide compounds, namely surfactin and iturin.
[0021] Preferably, the extraction step of the metabolite is as follows:
[0022] Step one: culturing bacillus amyloliquefaciens ZN-S18 in CM culture solution for 3 days;
[0023] Step two: centrifuging the bacterial solution and collecting supernatant;
[0024] Step three: filtering the supernatant obtained in step two through a 0.22-micron filter membrane to obtain sterile culture supernatant containing the metabolite;
[0025] Step four: confirming the metabolite by liquid chromatography-mass spectrometry analysis.
[0026] The main metabolic products of the Bacillus amyloliquefaciens ZN-S18 are surfactin and subtilin.
[0027] The application of the Bacillus amyloliquefaciens with the function of preventing and treating tomato diseases, the Bacillus amyloliquefaciens ZN-S18 can inhibit the mycelial growth of fusarium oxysporum f.sp lycopersici and f.sp fragariae, botrytis cinerea and botrytis cinerea, and the proliferation of fusarium solani, and can also inhibit the spore germination of fusarium oxysporum f.sp lycopersici and the sporulation of fusarium oxysporum and botrytis cinerea.
[0028] The application of the biocontrol agent, which has the prevention and treatment effect on tomato wilt and bacterial wilt, and can inhibit the sporulation of fusarium oxysporum and botrytis cinerea.
[0029] The application of the metabolic product, the metabolic product of the Bacillus amyloliquefaciens ZN-S18 can inhibit the mycelial growth and spore germination of fusarium oxysporum.
[0030] Compared with the prior art, the application provides the Bacillus amyloliquefaciens with the function of preventing and treating tomato diseases and the application thereof, and has the following beneficial effects:
[0031] 1. The Bacillus amyloliquefaciens ZN-S18 strain with the function of preventing and treating tomato diseases has a significant effect in the prevention and treatment of tomato wilt, and also has broad-spectrum antagonistic activity in inhibiting various pathogenic bacteria, including the mycelial growth of fusarium oxysporum f.sp lycopersici and f.sp fragariae, botrytis cinerea and botrytis cinerea, and the proliferation of fusarium solani, and can also significantly inhibit the spore germination of fusarium oxysporum f.sp lycopersici and the sporulation of fusarium oxysporum and botrytis cinerea; the microbial agent prepared from the Bacillus amyloliquefaciens ZN-S18 has good prevention and treatment effect on tomato wilt and bacterial wilt in tomato seedlings, and can also inhibit the sporulation of fusarium oxysporum and botrytis cinerea, and has a broad application prospect.
[0032] 2. The metabolic product of the Bacillus amyloliquefaciens ZN-S18 with the function of preventing and treating tomato diseases mainly includes two kinds of cyclic lipopeptide compounds, surfactin and subtilin, and both have clear disease resistance activity, can effectively inhibit the mycelial growth and spore germination of fusarium oxysporum, and show good biological control potential. DETAILED DESCRIPTION
[0033] Figure 1The endophyte isolated from the healthy tomato stem part has inhibitory activity on the Fusarium oxysporum f. sp. radicis-lycopersici.
[0034] Figure 2 The phylogenetic tree analysis diagram of Bacillus amyloliquefaciens ZN-S18 in the present application;
[0035] Figure 3 The inhibitory effect diagram of Bacillus amyloliquefaciens ZN-S18 in the present application in CM medium;
[0036] Figure 4 The phenomenon diagram of Bacillus amyloliquefaciens ZN-S18 in the present application in the culture solution;
[0037] Figure 5 The liquid chromatography-mass spectrometry analysis diagram of Bacillus amyloliquefaciens ZN-S18 in the present application in the culture solution;
[0038] Figure 6 The influence diagram of Bacillus amyloliquefaciens ZN-S18 on tomato seedlings in the present application;
[0039] Figure 7 The inhibition diagram of Bacillus amyloliquefaciens ZN-S18 on Pseudomonas solanacearum in the present application;
[0040] Figure 8 The inhibition diagram of Bacillus amyloliquefaciens ZN-S18 on the sporulation of Botrytis cinerea and Fusarium oxysporum in the present application. DETAILED DESCRIPTION
[0041] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.
[0042] Please refer to Figures 1-8 ;
[0043] Culture medium formula
[0044] Potato dextrose agar medium (PDA): potato 200 g, glucose 20 g, agar powder 15 g, distilled water 1 L, 121℃ sterilization for 15 min, natural pH.
[0045] Potato dextrose broth (PDB): potato 200 g, glucose 20 g, distilled water 1 L, 121℃ sterilization for 15 min, natural pH.
[0046] Complete Medium (CM): NaNO3 2 g, peptone 2.5 g, yeast extract 1 g, sucrose 30 g, K2HPO4 1 g, MgSO4.7H2O 0.5 g, KCl 0.5 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0047] Complete Medium (CM): NaNO3 2 g, peptone 2.5 g, yeast extract 1 g, sucrose 30 g, K2HPO4 1 g, MgSO4.7H2O 0.5 g, KCl 0.5 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0048] Minimal Medium (MM): NaNO3 2 g, sucrose 30 g, K2HPO4 1 g, MgSO4.7H2O 0.5 g, KCl 0.5 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0049] LB Medium: tryptone 10 g, yeast extract 5 g, NaCl 10 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0050] LB Medium: tryptone 10 g, yeast extract 5 g, NaCl 10 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0051] YMA Medium: yeast extract 4 g, malt extract 4 g, sucrose 4 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0052] CPG Medium: casein 1 g, tryptone 10 g, glucose 5 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0053] CPG Medium: casein 1 g, tryptone 10 g, glucose 5 g, agar powder 15 g, distilled water 1 L, sterilized at 121°C for 15 min, natural pH.
[0054] Example 1:
[0055] Isolation, purification, screening and identification of strains:
[0056] The stems of healthy tomato (Diana variety) plants were surface sterilized (treated with 70% alcohol, 3% sodium hypochlorite solution, and sterile water in sequence, and then air dried) and then ground into a homogenate with a homogenizer. The homogenate was diluted with sterile water to 10 -1 ,10 -2 ,10 -3 ,10 -4 concentration gradients, and 100 μL of each dilution was spread on LB medium. Each dilution gradient was set up in 5 replicates, and the plates were incubated at 28°C for 2 days. Different colonies that grew were picked and purified by the three-line method to obtain single colonies. The strains were numbered and stored at -80°C for long-term preservation.
[0057] To screen the most significant antagonistic strain, the Fusarium oxysporum (tomato transformation type) grown for 3 days was inoculated in the center of LB medium, and the endophytic strains isolated were inoculated on both sides of the plate. The plate was wrapped with tin paper and cultured at 25°C for 3 days, and the antagonistic effect was observed. According to the inhibition degree of Fusarium oxysporum mycelial growth, the endophytic strain with the most significant inhibition effect was screened.
[0058] The dominant strain ZN-S18 screened was inoculated in LB culture solution, and the bacterial cells were collected after being cultured at 28°C and 180 rpm for 1 day. The DNA was extracted, and the 16S rRNA gene was amplified by PCR using bacterial universal primers. The amplification product was subjected to sequence analysis, and the sequence was as follows:
[0059] TAGGGAAGAACAAGTGCCGTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAGAAAGCCA
[0060] CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGC
[0061] GTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCA
[0062] TTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGT
[0063] AGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAA
[0064] AGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGT
[0065] TAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCG
[0066] CAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGA
[0067] AGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTT
[0068] CGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTG
[0069] The sequencing results were analyzed by BLAST comparison in the NCBI database, and the bacterial universal marker gene rpoB was further amplified and sequenced, and the 16S rRNA sequence was combined to construct a phylogenetic tree. The results showed that the endophytic strain belongs to Bacillus amyloliquefaciens. Finally, it was named Bacillus amyloliquefaciens ZN-S18, and was preserved in the China General Microbiological Culture Collection Center (CGMCC) on September 9, 2025, with the preservation number of CGMCC No. 35860.
[0070] Example Two:
[0071] Optimal medium screening:
[0072] The confrontation culture of ZN-S18 and Fusarium oxysporum was carried out in CM, PDA and MM media, and the control was the confrontation culture of Fusarium oxysporum and Pseudomonas syringae or Escherichia coli. The results were observed after 3 days of culture at 25°C. The experiment showed that ZN-S18 had the most significant inhibitory effect in CM medium.
[0073] Example Three:
[0074] Effect of Bacillus amyloliquefaciens ZN-S18 culture solution on mycelial growth and spore germination of Fusarium oxysporum:
[0075] Bacillus amyloliquefaciens ZN-S18 was inoculated in CM culture solution and cultured at 28°C with 180 rpm shaking for 3 days. After the culture was completed, the bacterial bodies were removed by centrifugation at 13000 rpm, and the supernatant was filtered through a 0.22 μm filter membrane to obtain sterile culture solution containing metabolic products.
[0076] In the spore germination experiment, the sterile culture solution was mixed with the CM culture solution at a volume ratio of 1:1 as the treatment group culture solution; the control group culture solution was mixed by CM culture solution and sterile water at a ratio of 1:1. In the mycelial growth experiment, 1 mL of sterile culture solution was evenly coated on the surface of CM medium as the treatment medium.
[0077] The method for preparing the spores of Fusarium oxysporum is as follows: Fusarium oxysporum is inoculated into CM culture medium and cultured at 25°C for 3 days; then 5-6 pieces of the inoculum are taken and cultured in CM culture medium at 25°C with shaking at 180 rpm for 3 days, and the mycelium is collected and washed twice with sterile water. Then the mycelium is crushed with a spoon and uniformly spread on YMA culture medium, and induced to culture for 2-3 days under blue light to obtain spores. After the spores are collected, the concentration is adjusted to 10 5 spores / mL, inoculated into the culture medium of the treatment group and the control group, and cultured at 25°C with shaking at 180 rpm for 24 hours, and the spore germination is observed every 4 hours and the germination rate is calculated. The results show that the culture medium of Bacillus amyloliquefaciens ZN-S18 can significantly inhibit the germination of Fusarium oxysporum spores.
[0078] Further, the spores cultured for 24 hours are collected by centrifugation, washed twice with sterile water, and then re-inoculated into fresh CM culture medium and cultured at 25°C with shaking at 180 rpm for 12 hours. The results show that the spore germination rate does not recover, and part of the spores appear obvious swelling.
[0079] In addition, the mycelium of Fusarium oxysporum grown for 3 days is inoculated on the CM culture medium treated with the culture medium containing Bacillus amyloliquefaciens ZN-S18, and the results show that the mycelium growth of Fusarium oxysporum is significantly inhibited.
[0080] Example Four:
[0081] Detection of antibacterial substances in the culture medium:
[0082] The strain ZN-S18 is inoculated into CM culture medium and cultured at 28°C with shaking at 180 rpm for 3 days. Then the bacterial bodies are removed by centrifugation at 13000 rpm, and the sterile supernatant is obtained by filtration through a 0.22 μm filter membrane. After the supernatant is freeze-dried to remove water, 1 mL of methanol is added for dissolution. The obtained solution is subjected to liquid chromatography mass spectrometry analysis. The results show that the culture medium of ZN-S18 contains two types of cyclic lipopeptide compounds, surfactin and iturin;
[0083] Example Five:
[0084] Inhibition of Fusarium oxysporum pathogenicity by Bacillus amyloliquefaciens ZN-S18 in tomato seedlings:
[0085] ZN-S18 is inoculated into CM culture medium and cultured at 28°C with shaking at 180 rpm for 1 day, and then the bacterial bodies are collected by centrifugation and washed twice with sterile water. Then the bacterial suspension is diluted with sterile water to OD600=0.1. The spores of Fusarium oxysporum are collected according to the method of Example 3, and the concentration is adjusted to 10 5 spores / mL with sterile water.
[0086] Tomato seeds were surface sterilized (soaked in 70% alcohol, 3% sodium hypochlorite solution, and sterile water, and dried), and then germinated by using the wet cloth wrapping method. After 7 days, the germinated seedlings were transplanted into vermiculite substrate. After 24 hours, the seedlings were irrigated with Bacillus amyloliquefaciens ZN-S18 bacterial suspension; after 48 hours, the tomato seedlings were inoculated with Fusarium oxysporum spore suspension. The above process was repeated once a week.
[0087] The treated tomato seedlings were cultured in an artificial climate chamber (conditions: 12 hours light / 12 hours darkness, 25°C), and the results were observed after 21 days of continuous culture. The results showed that the tomato seedlings treated with Bacillus amyloliquefaciens ZN-S18 had significantly enhanced resistance to Fusarium oxysporum, effectively inhibiting the infection of the pathogenic bacteria.
[0088] Example Six:
[0089] R. solanacearum and Bacillus amyloliquefaciens ZN-S18 were inoculated into CPG culture medium and CM culture medium, respectively, and cultured at 28°C and 180 rpm for 1 day. Then the bacterial cells were collected by centrifugation and washed twice with sterile water, and the bacterial suspension was diluted with sterile water to adjust the OD600 of the two strains to 0.1.
[0090] In CM, CPG, and LB culture media, 5 μL of R. solanacearum and ZN-S18 were simultaneously inoculated, and the culture media were wrapped with tin foil. After 2 days of culture at 28°C, the results were observed. The results showed that Bacillus amyloliquefaciens ZN-S18 could significantly inhibit the growth of R. solanacearum in the three different culture media.
[0091] In addition, R. solanacearum with OD600=0.1 was inoculated into the culture medium obtained by mixing the culture medium of Bacillus amyloliquefaciens ZN-S18 and CPG culture medium at a volume ratio of 1:1 (preparation method same as Example 3), and the growth was observed after 12 hours of culture at 28°C and 180 rpm. The results showed that the addition of Bacillus amyloliquefaciens ZN-S18 culture medium could significantly inhibit the growth of R. solanacearum.
[0092] Further confrontation culture experiments of Botrytis cinerea were carried out in CM medium, and the results showed that Bacillus amyloliquefaciens ZN-S18 could effectively inhibit the growth of Botrytis cinerea mycelium. At the same time, Botrytis cinerea or Fusarium oxysporum was inoculated at one end of the CM medium, and ZN-S18 was inoculated at the other end, and the medium between the two was removed. After inoculation, the medium was sealed with sealing film and wrapped with tin paper, and the growth of the pathogenic fungal mycelium was observed after 3 days of culture at 25 DEG C. In addition, the fungal mycelium of Botrytis cinerea and Fusarium oxysporum was sampled by a puncher, placed in a centrifuge tube, 1 mL of sterile water was added, shaken to release spores, and the number of spores was counted. The results showed that Bacillus amyloliquefaciens ZN-S18 could produce volatile substances, which not only significantly inhibited the growth of Botrytis cinerea and Fusarium oxysporum mycelium, but also effectively inhibited the sporulation of the two pathogenic fungi.
[0093] The Bacillus amyloliquefaciens ZN-S18 strain with tomato disease control function has a significant effect in the prevention and treatment of tomato wilt, and also shows broad-spectrum antagonistic activity in inhibiting a variety of pathogenic fungi, including the mycelial growth of Fusarium oxysporum (tomato and strawberry specializations), Colletotrichum gloeosporioides, Botrytis cinerea, and the proliferation of Pseudomonas solanacearum. Bacillus amyloliquefaciens ZN-S18 can also significantly inhibit the spore germination of Fusarium oxysporum (tomato specialization) and the sporulation of Fusarium oxysporum and Botrytis cinerea. The microbial agent prepared from Bacillus amyloliquefaciens ZN-S18 shows good control effect on tomato wilt and bacterial wilt in tomato seedlings, and can inhibit the sporulation of Fusarium oxysporum and Botrytis cinerea, and has a broad application prospect. The metabolites of Bacillus amyloliquefaciens ZN-S18 with tomato disease control function mainly include two types of cyclic lipopeptide compounds, surfactin and subtilin, both of which have clear disease resistance activity and can effectively inhibit the mycelial growth and spore germination of Fusarium oxysporum, showing good biological control potential.
[0094] Although the embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and alterations can be made without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.
Claims
1. A type of Bacillus amyloliquefaciens with the function of preventing and controlling tomato diseases, characterized in that, Identified as Bacillus amyloliquefaciens ZN-S18, it was deposited on September 9, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 35860, located at Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen No. 1 Courtyard, Chaoyang District, Beijing.
2. The biocontrol agent according to claim 1, characterized in that, The biocontrol agent includes Bacillus amyloliquefaciens ZN-S18 as described in claim 1.
3. The biocontrol agent according to claim 2, characterized in that, The preparation method of the biocontrol agent includes the following steps: Step 1: Inoculate activated Bacillus amyloliquefaciens ZN-S18 into CM culture medium and incubate for 24 hours; Step 2: When the OD of the culture medium... 600 When the value reaches 1.0-1.5, a biocontrol agent is obtained.
4. A metabolite according to claim 1, characterized in that, The metabolite is the metabolite of Bacillus amyloliquefaciens ZN-S18 as described in claim 1.
5. A metabolite according to claim 4, characterized in that, The ZN-S18 metabolites include two types of cyclic lipopeptide compounds: surfactantin and iturin.
6. A metabolite according to claim 4, characterized in that, The extraction steps for the metabolites are as follows: Step 1: Incubate Bacillus amyloliquefaciens ZN-S18 in CM medium for 3 days; Step 2: Centrifuge the bacterial culture and collect the supernatant; Step 3: Filter the supernatant obtained in Step 2 through a 0.22μm filter membrane to obtain a sterile culture supernatant containing metabolites; Step 4: Confirm using liquid chromatography-mass spectrometry analysis: The main metabolites of Bacillus amyloliquefaciens ZN-S18 are surfactant and subtilisin.
7. The application of Bacillus amyloliquefaciens with the function of preventing and controlling tomato diseases according to claim 1, characterized in that: Bacillus amyloliquefaciens ZN-S18 can inhibit the mycelial growth of Fusarium oxysporum (tomato-specific and strawberry-specific), anthracnose fungus, and Botrytis cinerea, as well as the proliferation of Ralstonia solanacearum. Bacillus amyloliquefaciens ZN-S18 can also inhibit the spore germination of Fusarium oxysporum (tomato-specific) and inhibit the sporulation of Fusarium oxysporum and Botrytis cinerea.
8. The application of the biocontrol agent according to claim 2, characterized in that, It has a preventive effect against tomato wilt and bacterial wilt in tomato seedlings, and can inhibit sporulation of Fusarium oxysporum and Botrytis cinerea.
9. The application of a metabolite according to claim 4, characterized in that, Metabolites of Bacillus amyloliquefaciens ZN-S18 can inhibit the mycelial growth and spore germination of Fusarium oxysporum.