Bacillus velezensis CL02 and application thereof
By using Bacillus vesiculus CL02 and its fermentation broth and volatile gases, the problem of postharvest rot of chestnut fruits was solved, achieving a highly efficient biological control effect while maintaining fruit quality.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- CHANGLI INST OF POMOLOGY HEBEI ACADEMY OF AGRI & FORESTRY SCI
- Filing Date
- 2026-01-21
- Publication Date
- 2026-04-10
AI Technical Summary
Existing technologies are insufficient to effectively control post-harvest decay of chestnut fruits, chemical control methods pose environmental pollution risks, and there is a lack of efficient biological control methods.
By using Bacillus bellis CL02 and its fermentation broth and volatile gases, the growth of chestnut fruit rot pathogens is inhibited, thus preventing fruit rot and maintaining fruit quality.
It significantly inhibits the spread of various chestnut fruit rot pathogens, reduces the fruit rot rate, and maintains the quality of the fruit such as starch and soluble sugars, without affecting the vitamin C content.
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Figure CN121825822A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of plant disease control technology, specifically to Bacillus belye CL02 and its applications. Background Technology
[0002] Chestnut( Castanea mollissima Chestnuts (Brassica juncea) are a globally popular forestry crop due to their nutritional value, unique taste, and pleasant aroma. Chestnuts are rich in starch, protein, functional polysaccharides, essential fatty acids, vitamins, and minerals. Furthermore, chestnuts offer numerous health benefits, such as lowering cholesterol, preventing obesity and diabetes, fighting tumors, and improving the immune system. However, due to their high water content and rich nutrients, chestnuts are susceptible to pathogen infection during harvesting and storage, leading to rot and spoilage, severely impacting their quality and commercial value. Currently, approximately more than half of chestnut production losses are caused by rot and spoilage during storage and transportation. Several pathogens have been identified in chestnut fruit rot, including Penicillium (Brassica juncea). Penicillium spp. Alternaria ( ) Alternaria spp. ), Mucor ( Mucor spp. ), Rhizopus ( Rhizopus spp. ), Staphylococcus ( Botryosphaeria spp. ), Sclerotium genus ( Sclerotinia spp. Fusarium ( ) Fusarium spp. ), Microsporum ( Parvum spp. ) and Trichoderma ( Trichothecium spp. After infection by the pathogen, the contents of total soluble solids, soluble protein, starch, moisture and malondialdehyde in chestnut fruits all decreased significantly.
[0003] Currently, effectively controlling post-harvest rot remains a pressing issue for the chestnut industry. In production, prevention and control of chestnut fruit rot primarily involve reducing mechanical damage during harvesting, post-harvest storage, and transportation, as well as using chemical fungicides before harvest to minimize pathogen infection. However, these measures are insufficient to control the occurrence of chestnut fruit rot, and chemical control can easily cause environmental pollution and food safety issues. Therefore, researching and promoting new environmentally friendly biological control methods has irreplaceable advantages. Microbial agents have become a research hotspot due to their safety, high efficiency, economy, and environmental friendliness. However, current research lacks microbial agents that can effectively control various chestnut fruit rot pathogens. Summary of the Invention
[0004] In view of the above technical problems, the present invention provides Bacillus belyssus CL02 and its applications.
[0005] The present invention adopts the following technical solution: The application provides bacillus velezensis CL02, which is classified as bacillus velezensis Bacillus velezensis , and is preserved in the China General Microbiological Culture Collection Center on July 14, 2025, and has a preservation number of CGMCC No. 35215.
[0006] The application aims at the problems of environmental pollution of chemical agents and lack of existing biocontrol agents in the prevention and treatment of postharvest fruit rot of Chinese chestnut, and a bacillus velezensis CL02 is obtained by screening from the rhizosphere soil of Chinese chestnut. Diaporthe ere s, The bacillus velezensis CL02, fermentation broth and volatile gas thereof have significant inhibitory activity on a plurality of Chinese chestnut fruit rot pathogens including Diaporthe phaseolorum var. succulenta D. ere s , can cause abnormality of mycelium of the pathogens, and effectively inhibit the expansion of fruit diseases. Meanwhile, the CL02 treatment does not affect the main quality indexes such as starch and soluble sugar of Chinese chestnut during storage, and can maintain the VC content, so that the fruit quality is guaranteed while the prevention and treatment effect is ensured, and new strain resources and technical approaches are provided for the development of safe and efficient postharvest biological rot prevention agents of Chinese chestnut.
[0007] The application also provides application of the bacillus velezensis CL02 in prevention and treatment of Chinese chestnut fruit rot.
[0008] Further, the pathogenic bacteria of the Chinese chestnut fruit rot include Diaporthe phaseolorum var. succulenta, Alternaria alternata, Mucor circinelloides, Rhizopus stolonifer and Fusarium proliferatum.
[0009] The application also provides application of the bacillus velezensis CL02 in preparation of a product for Chinese chestnut fruit rot, and the active ingredient of the product is the bacillus velezensis CL02.
[0010] Further, the active ingredient of the product is fermentation broth and / or volatile gas of the bacillus velezensis CL02.
[0011] Further, the product is a biological pesticide or a rot prevention agent.
[0012] Further, the product is a solution type.
[0013] Further, the solvent of the product is water or glycerol.
[0014] The application has the following beneficial effects: The experiments prove that the bacillus velezensis CL02, the fermentation liquor and the volatile gas thereof can effectively inhibit the mycelium expansion of the chestnut fruit rot pathogen Diaporthe phaseolorum, cause the mycelium swelling and deformation of the pathogenic bacteria, and significantly inhibit the expansion of the chestnut fruit rot pathogen Diaporthe phaseolorum in the kernel. The strain CL02 has significant bacteriostatic activity on pathogenic bacteria of other genera of chestnut fruit rot, chestnut blight, cherry gray mold and other common fungi. The strain CL02 has no damage to the quality of the chestnut fruit during the storage period, including no significant influence on starch, soluble sugar, soluble protein, soluble solids and water content, and can maintain the VC content of the chestnut. In addition, the fermentation liquor, sterile fermentation liquor and volatile gas of CL02 can significantly reduce the natural rot rate of the fruit during the low-temperature storage period, and have great application potential. BRIEF DESCRIPTION OF DRAWINGS
[0015] Figure 1 The inhibition effect diagram of the CL02 strain on D. ere s .
[0016] Figure 2 The phylogenetic analysis diagram of the strain CL02.
[0017] Figure 3 The inhibition rate diagram of the CL02 bacterial body, volatile gas (VOCs) and sterile fermentation liquor (CFS) on the growth of D. ere s .
[0018] Figure 4 The mycelium swelling and deformation diagram induced by the CL02. D. ere s
[0019] Figure 5 The bacteriostatic spectrum diagram of the CL02 strain.
[0020] Figure 6 The live body prevention and cure effect diagram of the CL02 bacterial body, volatile gas (VOCs) and sterile fermentation liquor (CFS) on the chestnut fruit rot.
[0021] Figure 7 The inhibition diagram of the CL02 bacterial body, volatile gas (VOCs) and sterile fermentation liquor (CFS) on the growth of the chestnut fruit rot pathogen D. ere s .
[0022] Figure 8 The statistical diagram of the influence of the CL02 bacterial body, volatile gas (VOCs) and sterile fermentation liquor (CFS) treatment on the chestnut rot rate.
[0023] Figure 9 The influence diagram of the CL02 on the natural rot rate and storage quality of the chestnut during the low-temperature storage period, wherein A is the starch content statistical diagram, B is the soluble sugar content statistical diagram, C is the soluble protein content statistical diagram, D is the soluble solid content statistical diagram, E is the water content statistical diagram, and F is the VC content statistical diagram. Detailed Implementation
[0024] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments, but this should not be construed as limiting the invention. Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following embodiments are commercially available unless otherwise specified.
[0025] Chestnut materials used for testing: The chestnut fruits used for testing were collected from the chestnut germplasm resource nursery of the Science and Technology Innovation Base of Changli Fruit Tree Research Institute, Hebei Academy of Agricultural and Forestry Sciences. The varieties were named Yanshan Zaofeng and were stored in the cold storage of Changli Fruit Tree Research Institute, Hebei Academy of Agricultural and Forestry Sciences (3±0.5℃).
[0026] Test culture media: Potato glucose agar (PDA) medium (200g potato, 20g glucose, 15g agar, 1L distilled water); NA medium (3g beef extract, 5g peptone, 2.5g glucose, 1L water; 15g agar added to solid culture medium before sterilization).
[0027] Example 1: Identification of Bacillus belyssus CL02.
[0028] I. Purification of biocontrol bacteria.
[0029] In April 2025, rhizosphere soil samples from chestnut trees were collected at the chestnut germplasm resource nursery of the Changli Fruit Tree Research Institute, Hebei Academy of Agricultural and Forestry Sciences. Bacteria in the soil were isolated using the dilution plate method. 10g of soil sample was thoroughly ground, then transferred to 90mL of sterile water and mixed. The mixture was incubated at 28℃ and 200rpm for 3 hours. The soil suspension was then serially diluted to 10... -3 10 -4 10 -5 The concentration of the diluted solution was measured, and 100 μL of the diluted solution was evenly spread on NA solid medium. The medium was incubated upside down at 28°C for 24 h. Colonies of different sizes and shapes were picked and streaked onto new NA medium plates for purification.
[0030] II. Screening of biocontrol bacteria.
[0031] The inhibitory activity of candidate biocontrol strains against the chestnut fruit rot fungus *Citrus sweet cherry* was determined using the plate confrontation method. This target fungus has been disclosed in the literature “Xu, H.; Zhao, W.; Guo, Y.; Cui, J.; Niu, G.; Zhang, S.; Li, Y.; Li, L.; Jiao, R.; Wang, X.; Wang, G.; He, L. Identification of Fungal Pathogens of Chinese Chestnut Fruit Rot and Analysis of Resistance Differences among Major Cultivars. Microorganisms 2026, 14, 113”. The specific procedures are as follows: Take a 5mm mycelial disc from the edge of a PDA plate containing the chestnut fruit rot fungus *Saccharum spp.*, cultured for 7 days, and transfer it to the center of a new PDA plate. Pick a single colony of the purified biocontrol strain, mix it in 100μL of sterile water, spread it on a new NA plate, and incubate at 28℃ for 24 hours. Take four 5mm mycelial discs from the culture and transfer them 2.5cm from the center of the PDA plate. A PDA plate containing only the chestnut pathogen serves as a control. Incubate the plates at 28℃ for 5 days. Each treatment is repeated three times, and the experiment is repeated twice. Inhibition rate (%) = (colon diameter of control group − colony diameter of treatment group) / colony diameter of control group × 100%.
[0032] Experimental results are as follows Figure 1 As shown, CL02 significantly inhibited the growth of chestnut fruit rot pathogens. D. ere s Mycelial growth.
[0033] III. Identification of biocontrol bacteria.
[0034] Total DNA was extracted from biocontrol bacteria and subjected to 16S rRNA and... gyrB PCR amplification of the gene. PCR amplification system: (50μL system) 25μL 2×Taq PCR mixture, 1μL forward primer, 1μL reverse primer, 1μL template, 22μL ddH2O. PCR program: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 1 min, 55℃ annealing for 1 min, 72℃ extension for 2 min, 34 cycles; 72℃ extension for 10 min. Amplified products were sequenced by Qingke Biotechnology (Beijing, China). The sequences were analyzed using BLAST and submitted to the NCBI database. Further sequence alignment and homology analysis were performed using ClustlX and MEGA7, respectively, to construct a phylogenetic tree.
[0035] Experimental results are as follows Figure 2 As shown, multiple genes (16S and gyrbCombined phylogenetic analysis identified strain CL02 as... Bacillus velezensis .
[0036] Example 2: Antibacterial analysis.
[0037] I. Antimicrobial analysis of sterile fermentation filtrate and volatile compounds of Bacillus belyss CL02.
[0038] 1. The growth rate method was used to determine the effect of CL02 sterile fermentation filtrate on the growth of bacteria. D. ere s Inhibitory effect: Single colonies of strain CL02 were transferred to 2 mL of NA liquid medium and incubated at 28°C and 220 rpm for 24 h with shaking. The colonies were then transferred to 100 mL of fresh NA liquid medium and incubated at 28°C and 220 rpm for another 24 h to obtain the fermentation broth. The fermentation broth was collected in 50 mL centrifuge tubes, centrifuged at 12000 rpm for 20 min, and the supernatant was filtered twice through a 0.22 μm sterile filter membrane. The sterile fermentation broth was added to PDA medium at 50°C at volume fractions of 1%, 5%, 10%, and 20%, mixed thoroughly, and then poured into plates. PDA solid medium plates containing no sterile fermentation filtrate (0%) served as a control. Each treatment was repeated three times. After the medium solidified, a 5 mm inoculum was planted in the center. D. ere s Mycelial cakes. Plates were incubated at 28°C for 5 days, and colony diameters were measured to calculate the inhibition rate.
[0039] 2. The effect of volatile compounds generated by ClO2 on the [unclear - possibly related to environmental factors] was determined using a two-plate method. D. ere s Inhibitory effect: Single colonies of strain CL02 were transferred into 2 mL of NA liquid medium and incubated at 28°C and 220 rpm for 24 h. 100 μL of the fermentation broth was then evenly spread onto an NA plate, and a 5 mm diameter section was added. D. ere s Place the tablet in the center of the PDA plate and seal the two plates face to face. The control group was inoculated. D. ere s PDA plates and NA plates without CL02 inoculation were sealed together. Each treatment was repeated in triplicate, and the experiment was repeated twice. The plates were incubated at 28°C for 5 days, and the colony diameter was measured to calculate the inhibition rate.
[0040] Experimental results are as follows Figure 3 As shown, CL02 cells have a significant effect on... D. ere s The mycelial inhibition rate reached 81.98%; volatile gas treatment and 20% (v / v) sterile fermentation filtrate treatment had an effect on... D. ere s The inhibition rates were 63.58% and 69.26%, respectively.
[0041] II. Observation of hyphal morphology and hyphal microstructure.
[0042] Cut 1cm from the edge of the inhibition zone. 2 of D. ere sMycelium, and compared with those grown on normal culture medium D. ere s The mycelia were compared. Morphological changes in the mycelia were observed using an electron microscope.
[0043] Experimental results are as follows Figure 4 As shown, CL02 can induce D. ere s The hyphae are swollen and deformed.
[0044] III. Evaluation of antibacterial spectrum.
[0045] The inhibitory effect of CL02 on six common plant pathogenic fungi was analyzed using the plate confrontation method, including other pathogens of chestnut fruit rot fungus and Alternaria alternata. Alternaria alternate Mucor ( ), Mucor circinelloides ), Rhizopus spp. ( Rhizopus stolonifer Fusarium ( ), Fusarium proliferatum ), and chestnut blight pathogen ( Cryphonectria parasitica ), cherry gray mold ( Botrytis cinerea Incubate at 28℃ for 6 days, and measure colony diameter to calculate inhibition rate. The Alternaria alternata (… A. alternate Mucor ( ), M. circinelloides ), Rhizopus spp. ( R. stolonifer Fusarium ( ), F. proliferatum The fungus *Phytophthora indicum* (Chestnut blight pathogen) was first identified in the literature “Xu, H.; Zhao, W.; Guo, Y.; Cui, J.; Niu, G.; Zhang, S.; Li, Y.; Li, L.; Jiao, R.; Wang, X.; Wang, G.; He, L. Identification of Fungal Pathogens of Chinese Chestnut Fruit Rot and Analysis of Resistance Differences among Major Cultivars. Microorganisms 2026, 14, 113”. C. parasiticaZhao L, Wei X, Chen F, Yuan L, Chen B, Li R. N6-methyladenosine RNA methyltransferase CpMTA1 mediates CpAphA mRNA stability through a YTHDF1 -dependent m6A modification in the chestnut blight fungus. PLoS Pathog. 2024 Aug 19;20(8):e1012476. doi: 10.1371 / journal.ppat.1012476. in the literature. The cherry gray mold fungus (Cryphonectria parasitica) was disclosed in “Cui J, Xu HJ, Jia XH, et al. Isolation and identification of main postharvest fungal diseases of sweet cherry and analysis of pathogenicity [J]. Journal of Nuclear Agriculture, 2024, 38(10): 1968-1975.” in the literature. Botrytis cinerea
[0046] The experimental results are shown in Figure 5 , CL02 can significantly inhibit the growth of other pathogens of chestnut fruit rot fungus Alternaria alternata, Mucor circinelloides, Rhizopus stolonifer, Fusarium proliferatum, chestnut blight fungus and cherry gray mold fungus.
[0047] Example 3: In vivo antibacterial activity of chestnut.
[0048] I. In vivo control effect of chestnut fruit rot
[0049] The effect of antagonistic bacteria strain CL02 on the prevention and control of chestnut fruit rot was determined by chestnut stab inoculation test. The chestnut was soaked in 2% sodium hypochlorite solution for 3 min, washed with sterile water, and then dried with sterile filter paper. Then it was transferred to a covered polyethylene (PE) fresh-keeping box (31.5*23.5*11.5cm 3 ) with an OD 600 of 1.0, 20% sterile fermentation filtrate was sprayed on the surface of the chestnut until the droplets flowed down, and then naturally dried. The process was repeated 3 times. Sterile water was sprayed as a control. To test the in vivo effect of CL02 volatile gas, a 5-dish culture of CL02 was placed in the fresh-keeping box containing the chestnut for 24 h, and a sterile NA plate was placed as a control without direct contact with the chestnut. A sterile needle was used to make a wound on each chestnut, and the wound was inoculated with D. ere s A 5mm mycelial cake was prepared, with sterile PDA medium inoculated as a control. The container was covered with a food storage container and sprayed with sterile water to maintain humidity above 90% RH. All treatments were cultured at 25℃ under alternating light and dark conditions (12h / 12h) for 14 days. The diameter of the lesions was measured, and the development of chestnut fruit rot in different treatments was statistically analyzed.
[0050] Experimental results are as follows Figure 6 and Figure 7 As shown, the in vivo control efficacy of strain CL02 against chestnut fruit rot indicates that CL02 bacterial suspension, sterile fermentation broth, and volatile gases can all significantly inhibit [the disease]. D. ere s The inhibition rates of chestnut rot caused by these factors were 79.70%, 67.26%, and 70.25%, respectively.
[0051] II. The effect of CL02 treatment on the natural decay of chestnuts during storage.
[0052] Select 'Yanshan Zaofeng' chestnuts of uniform size, free from mechanical damage and pests, soak them in a 2% sodium hypochlorite solution for 3 minutes, rinse them with sterile water, and then blot dry with sterile filter paper. The fruits are then treated as follows: (1) Bacterial treatment: Prepare CL02 fermentation broth, collect the bacteria by centrifugation at 12000 rpm, resuspend them in sterile water, repeat the centrifugation and resuspending process twice, and finally dissolve the bacteria in sterile water and dilute to OD. 600 =1. Fruit soaked in OD 600 (1) After soaking in the bacterial solution for 30 minutes, the fruit was naturally dried and placed in a PE preservation bag; (2) Aseptic fermentation filtrate treatment: The fruit was soaked in the aseptic fermentation filtrate for 30 minutes and then naturally dried and placed in a PE preservation bag; (3) Volatile gas treatment: The fruit was spread flat in a sieve with a diameter of 30 cm and a mesh size of 5. 100 μL of fermentation liquid was evenly spread on NA plates in advance and cultured at 28℃ for 24 h. 10 CL02 culture plates were placed under each sieve without direct contact with the chestnuts. The plates and sieves were placed in a PE preservation bag. Fresh CL02 culture plates were replaced every 5 days; (4) Control treatment: The fruit was not treated and placed in a PE preservation bag. 1 kg of fruit was placed in each bag for the above treatments. Each treatment was repeated 3 times and stored in a cold storage at (3±0.5)℃. When the fruit temperature was the same as the storage temperature (2 h), the bag opening was sealed tightly. Chestnut fruits from each treatment were sampled at (3±0.5)℃ for 0, 15, 30, 45, and 60 days of storage. Each sample was collected in one preservation bag. Statistical analysis of fruit rot rate was performed. Rot rate (%) = (Number of rotten fruits / Total number of fruits) × 100%.
[0053] Experimental results are as follows Figure 8As shown, during the low-temperature storage period, CL02 bacterial solution, fermentation broth, and volatile gases can all significantly inhibit the natural decay rate of chestnuts; there was no significant difference in decay rate among the three treatments in the first 45 days; after 60 days, the decay rate of chestnuts treated with bacterial solution and volatile gases was significantly lower than that of chestnuts treated with fermentation broth, and was also significantly lower than that of the control.
[0054] III. The effect of CL02 on the quality of chestnuts during storage.
[0055] Select 'Yanshan Zaofeng' chestnuts of uniform size, free from mechanical damage and pests, soak them in a 2% sodium hypochlorite solution for 3 minutes, rinse them thoroughly with sterile water, and blot dry with sterile filter paper. Then place them in CL02 bacterial solution (OD2002). 600 =1) Soak in water for 30 minutes, air dry naturally, and pack into PE preservation bags, with each bag containing 1 kg. Each treatment is repeated 3 times, with sterile water soaking as a control. The treated fruits are stored in a cold storage at a temperature of (3±0.5)℃. When the fruit temperature is the same as the storage temperature, the opening of the preservation bag is sealed tightly. Samples are taken at 0, 15, 30, 45, and 60 days of storage, with each sample consisting of one preservation bag. The fruits are placed at 20℃ for 24 hours to equilibrate and determine physicochemical indicators such as starch, soluble sugar, soluble protein, soluble solids, water content, and vitamin C content.
[0056] Starch content was determined by acid hydrolysis; soluble sugar content was determined by anthrone-sulfuric acid colorimetric method; vitamin C (VC) content was determined by 2,6-dichlorophenolindophenol titration method; soluble protein content was determined by Coomassie Brilliant Blue G-250 staining method; soluble solids content was determined by refractometer method using PAL-1 digital display saccharimeter; moisture content was determined according to GB / T5009.3-2003 "Determination of Moisture in Food".
[0057] Experimental results are as follows Figure 9 As shown, during the low-temperature storage period, there were no significant differences in starch, soluble sugar, soluble protein, soluble solids, and water content of chestnuts treated with CL02 bacterial solution compared to the control. However, after 15 days of storage, the vitamin C content of chestnuts treated with CL02 bacterial solution was significantly increased compared to the control, indicating that CL02 has no effect on the starch, soluble sugar, soluble protein, soluble solids, and water content of chestnuts, and can maintain the vitamin C content.
[0058] It should be noted that when numerical ranges are mentioned in the claims of this invention, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. To avoid redundancy, the present invention describes preferred embodiments.
[0059] While the preferred embodiments of the application have been described, additional variations and modifications can be made to these embodiments by those skilled in the art once they have the benefit of the present disclosure without departing from the spirit and scope of the application. Accordingly, it is intended that such additions and modifications be included within the scope of the application. It is the following claims, including any amendments thereto, which define the scope of the application.
[0060] Obviously, numerous modifications and variations of the present application are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims, and their equivalents, the application can be practiced otherwise than as specifically described.
Claims
1. Bacillus velezensis CL02, characterized in that, The classification name of the bacillus velezensis CL02 is bacillus velezensis Bacillus velezensis , and it is preserved in the China General Microbiological Culture Collection Center on July 14, 2025, with the preservation number of CGMCC No.35215.
2. The Bacillus velezensis CL02 of claim 1 is used for preventing and treating chestnut fruit rot.
3. The use of Bacillus velezensis CL02 according to claim 2 for preventing and treating Chinese chestnut fruit rot, characterized in that, The pathogenic bacteria of the chestnut fruit rot include Diaporthe phaseolorum, Alternaria alternata, Mucor circinelloides, Rhizopus stolonifer and Fusarium proliferatum.
4. Use of Bacillus velezensis CL02 according to claim 1 for the preparation of a product for the control of the chestnut fruit rot, characterized in that, The active ingredient of the product is the Bacillus velezensis CL02.
5. The use of B. velezensis CL02 according to claim 4 for the preparation of a product for the control of the chestnut fruit rot, characterized in that, The active ingredient of the product is the fermentation broth and / or volatile gas of the Bacillus velezensis CL02.
6. The use of B. velezensis CL02 according to claim 4 for the preparation of a product for the control of the chestnut fruit rot, characterized in that, The product is a biological pesticide or preservative.
7. The use of B. velezensis CL02 according to claim 6 for the preparation of a product for the control of the chestnut fruit rot, characterized in that, The product is in the form of a solution.
8. The use of B. velezensis CL02 according to claim 7 for the preparation of a product for the control of the chestnut fruit rot, characterized in that, The solvent of the product is water or glycerol.