Bacillus velezensis microbial inoculum for preventing and treating fusarium wilt and application of bacillus velezensis microbial inoculum

Abstract

The invention discloses a bacillus velezensis microbial agent for preventing and treating fusarium wilt and application thereof, relates to the technical field of fusarium wilt prevention and treatment, and solves the technical problems of pesticide residues and environmental pollution caused by adopting chemical pesticides to prevent and control fusarium wilt in the prior art. The bacillus velezensis microbial inoculum for preventing and treating fusarium wilt is bacillus velezensis fermentation liquor prepared by taking bacillus velezensis as a strain and carrying out seed activation and at least three-stage culture. The bacillus velezensis microbial agent for preventing and controlling the fusarium wilt is used for tomato planting, effective prevention and control of the fusarium wilt, activation of soil nutrients and growth promotion of crop root systems can be achieved in the seedling raising period, nutrient absorption of crops is enhanced after diseases are effectively prevented and controlled, and yield loss of the crops caused by the diseases is further reduced.

Description

Technical Field

[0001] The invention relates to the technical field of wilt prevention and treatment, in particular to a Bacillus Velezii agent for preventing and treating wilt and application thereof. Background Art

[0002] Fusarium oxysporum Fusarium oxysporum The tomato wilt disease caused by the disease can infect the roots of tomatoes and reproduce in the vascular bundles, causing the vascular bundles to be blocked, leading to the wilting and death of the plants. It has the characteristics of large damage and loss and is difficult to prevent and control. It can also harm a variety of economic crops such as melons, solanums, and beans. At present, chemical pesticides such as fludioxonil, benomyl, and prochloraz are mainly used to prevent and control tomato wilt disease. However, chemical pesticide treatment can easily cause environmental pollution and pesticide residues, which seriously endangers human and environmental health. Summary of the invention

[0003] The invention aims to provide a Bacillus Velezii agent for preventing and controlling wilt disease and application thereof, which solves the technical problems of pesticide residues and environmental pollution in the prior art when chemical pesticides are used to prevent and control wilt disease.

[0004] To achieve the above object, the present invention provides the following technical solutions: The first technical scheme of the present invention is to provide a Bacillus Velez subtilis agent for preventing and treating wilt disease. The agent is a Bacillus Velez subtilis fermentation liquid prepared by using Bacillus Velez subtilis YWF-075 as the strain and after seed activation and at least two-stage cultivation.

[0005] The second technical solution of the present invention is to provide the use of Bacillus Velez subtilis agent in the preparation of a preparation for preventing and treating diseases caused by tomato wilt pathogens.

[0006] The third technical solution of the present invention is to provide the use of the Bacillus Velez subtilis agent in the preparation of a seedling culture medium.

[0007] The fourth technical solution of the present invention is to provide the use of Bacillus Velez subtilis inoculant in the preparation of biological fertilizer.

[0008] The Velez subtilis agent for preventing and treating wilt disease and its application have at least the following beneficial technical effects: The Bacillus Velez microbial agent for preventing and controlling Fusarium wilt of the present invention is a fermented liquid of Bacillus Velez obtained by using Bacillus Velez as the strain and after seed activation and at least two-stage cultivation. When the fermented liquid is used in the planting of Solanaceae crops such as tomatoes, it can achieve effective prevention and control of Fusarium wilt, activation of soil nutrients and promotion of crop root growth in the seedling stage, enhance crop nutrient absorption after effective prevention and control of diseases, and further reduce crop yield losses caused by diseases. On the other hand, since the Bacillus Velez subtilis agent for preventing and controlling Fusarium wilt of the present invention can effectively prevent and control soil-borne diseases of tomatoes and other Solanaceae crops such as Fusarium wilt, it can greatly reduce the application amount of chemical pesticides, reduce environmental pollution, improve crop safety, and maintain the balance of soil microbial communities. That is, the Bacillus Velez subtilis agent for preventing and controlling Fusarium wilt of the present invention and its application solve the technical problems of the prior art of using chemical pesticides to prevent and control Fusarium wilt of Solanaceae crops such as tomatoes, resulting in pesticide residues and environmental pollution. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 This is a graph showing the antibacterial activity test results of the strains screened in Example 1 against wilt pathogens; Figure 2 This is a graph showing the antibacterial effect of the fermentation broth of Bacillus velezensis YWF-075 on Fusarium wilt. DETAILED DESCRIPTION

[0010] To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0011] A Bacillus Velez microbial agent for preventing and treating wilt disease, wherein the microbial agent is a Bacillus Velez fermentation liquid prepared by seed activation and at least two-stage culture using Bacillus Velez YWF-075 as the strain; wherein the number of effective live bacteria in the Bacillus Velez fermentation liquid is not less than 5×10 9 cfu / ml.

[0012] The classification name of the Bacillus Velez YWF-075 of the present invention is: Bacillus Velez ( Bacillus velezensis )YWF-075, the deposit registration number is CCTCC NO: M 20242105, the deposit date is September 27, 2024, the status is: alive, the depository unit is China Center for Type Culture Collection, and the deposit address is Wuhan University, China.

[0013] The invention discloses an application of the Bacillus Velezii agent for preventing and controlling Fusarium wilt in the preparation of a preparation for preventing and controlling diseases caused by tomato Fusarium wilt.

[0014] The bacterial agent is applied to tomatoes by one or more methods including root irrigation, pouring and hole application.

[0015] The bacterial agent is prepared into a suspension and diluted 100 to 300 times for application.

[0016] The application of the Bacillus Velez bacteria agent for preventing and controlling Fusarium wilt in the preparation of a seedling culture medium is used for seedling cultivation of Solanaceae crops.

[0017] The bacterial agent and the seedling culture medium are mixed in a weight ratio of 8 to 10:1000 and mixed evenly.

[0018] The invention discloses an application of the Bacillus Velezii agent for preventing and controlling wilt disease in the preparation of biological fertilizer.

[0019] The bacterial agent and the organic fertilizer are mixed in a weight ratio of 5 to 8:1000, mixed evenly, and used for fertilizing Solanaceae crops.

[0020] The following is attached to the instruction manual Figure 1 and 2 And Examples 1 to 4 describe in detail the Bacillus Velezii agent for preventing and controlling Fusarium wilt of the present invention and its application.

[0021] Example 1

[0022] This example illustrates the indoor activity determination of Bacillus velezensis YWF-075 of the present invention.

[0023] Antagonistic strains were obtained by the plate confrontation method. A 5 mm diameter puncher was used to take a block of Fusarium wilt pathogen hyphae and inoculated it in the center of the PDA culture dish. The cross method was used to dip the strain with an inoculation needle 3 cm away from the center of the plate. The treatment inoculated with Fusarium wilt pathogens was used as the control. The plate was placed in a 28 ℃ constant temperature incubator and cultured for 7 days until the pathogen hyphae on the control plate grew to the edge of the culture medium. Each treatment was repeated 3 times, and the strains with obvious inhibition zones were selected for antibacterial activity determination. The test results are shown in the attached Figure 1 shown.

[0024] Antagonistic activity test of indoor fermentation liquid: hole method was used: the wilt pathogen was inoculated in the center of the PDA culture dish, and a 0.5 cm hole punch was used to punch 3 cm from the center to remove the culture medium. 40 μL of YWF-075 fermentation liquid was taken with a pipette and placed in the hole. The control group was only inoculated with pathogens. After the plate was placed in a 28 ℃ constant temperature incubator for 5 days, the fermentation liquid of the strain was observed to see whether it had an inhibitory effect on the wilt pathogen, and the inhibition rate was calculated. Each treatment was repeated 3 times. The test results are shown in the attached figure. Figure 2 Through the antagonistic activity test, it was found that the Bacillus spore preparation YWF-075 had a strong inhibitory effect on the growth of wilt pathogens.

[0025] Example 2

[0026] This embodiment provides a spore-forming fungus agent and a method for directly applying the agent as the fungus agent.

[0027] In 2023, a plot test was conducted in the company's greenhouse. The test was carried out in plots with severe wilt disease in 2022. The bacterial agent was prepared into a suspension and diluted 100, 200, and 300 times at the early stage of tomato flowering, with an interval of 1 week each time. The treatments were 200-fold dilution of the Bacillus YWF-075 suspension preparation and 3% polyoxin, with clean water as the blank control. Each treatment was repeated 3 times, and the plots were randomly arranged. There were 48 plants in each treatment, and the management was the same as that of general fields. As can be seen from Table 1, in the plot test, both the Bacillus sporogenes YWF-075 preparation and 3% polyoxin can reduce the incidence of Fusarium wilt. Considering the preparation cost and control effect, the Bacillus sporogenes YWF-075 preparation diluted 200 times is the best choice, indicating that the Bacillus sporogenes YWF-075 preparation has a good application prospect for the control of Fusarium wilt.

[0028] Example 3

[0029] This embodiment provides a spore-forming fungus agent and a method for using the agent in combination with a matrix to prepare a functional seedling-growing matrix.

[0030] The bacterial agent of this embodiment is made by fermenting Bacillus velez YWF-075, and the effective viable bacteria count is not less than 5×10 9 cfu / ml of fermentation broth.

[0031] The spore-forming microbial agent of this embodiment can be combined with a matrix to prepare a functional seedling matrix for use in preventing and controlling diseases during the seedling stage, which can effectively reduce the outbreak of diseases after crop transplanting.

[0032] The specific method of use is as follows: mix the microbial agent with the seedling substrate (peat: vermiculite: perlite = 3:1:1) at a weight ratio of 8 to 10:1000, mix evenly and use it for seedling cultivation of crops, and cultivate seedlings according to conventional seedling cultivation methods. The crop seedlings obtained by this method already have disease-resistant bacteria in the root substrate soil and plants when transplanted, and can effectively prevent and control wilt disease after transplanting. After transplanting, the microbial agent can be used for 1 to 2 root irrigations or waterings in the early flowering period. Combined with Table 1, it can be seen that the microbial agent and the seedling substrate are mixed at a weight ratio of 8:1000 and used for crop seedling cultivation. Root irrigation once 7 days after transplanting can improve the prevention and control effect by more than 10%.

[0033] Example 4

[0034] This embodiment provides a spore-forming fungus agent and a method for using the agent in combination with an organic fertilizer to prepare a functional biological organic fertilizer.

[0035] The bacterial agent of this embodiment is made by fermenting Bacillus velez YWF-075, and the effective viable bacteria count is not less than 5×10 9 cfu / ml of fermentation broth.

[0036] The spore-forming microbial agent of this embodiment can be combined with the cloud leaf oil cake organic fertilizer to prepare a functional biological organic fertilizer for use. It can enter the soil when the base fertilizer is applied, and can be used to prevent and control diseases during the crop transplanting stage, which can effectively reduce the outbreak of diseases after crop transplanting.

[0037] The specific method of use is as follows: mix the microbial agent and the organic fertilizer in a weight ratio of 5 to 8:1000, mix evenly and use as base fertilizer when transplanting crops.

[0038] The specific method is: according to the fertilizer requirements of crops, the biological organic fertilizer in this embodiment is used as a base fertilizer when the crops are transplanted. Conventional base fertilizer application methods such as hole application and furrow application can be adopted. After application, the crops are transplanted and watered in time to promote the rapid germination of spores and spores in the biological organic fertilizer in a humid environment to form biological control. Other measures can be carried out according to conventional farming operations. According to this method, when crops are planted, colonization of disease-resistant bacteria can be formed in the soil around the root system during transplanting, and effective control of wilt can be formed after transplanting. Microbial agents can be used to irrigate the roots or pour them 1 to 2 times in the early flowering period after transplanting, which can improve the control effect by about 20%. As can be seen from Tables 1 and 3, the bacterial agent and organic fertilizer are mixed in a weight ratio of 6:1000 as base fertilizer, and applied once in the early flowering period after crop transplanting, and the control effect is improved by about 20%.

[0039] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A Bacillus Velezii agent for preventing and treating fusarium wilt, characterized in that: The microbial agent is a fermentation liquid of Bacillus velez YWF-075 which is prepared by seed activation and at least two-stage cultivation.

2. The Bacillus Velezii agent for preventing and treating Fusarium wilt as claimed in claim 1, characterized in that: The number of effective living bacteria in the fermentation broth of Bacillus Velez is not less than 5×10 9 cfu / ml.

3. Use of the Bacillus Velez subtilis agent for preventing and controlling Fusarium wilt as claimed in claim 1 in the preparation of a preparation for preventing and controlling diseases caused by tomato Fusarium wilt.

4. Use of the Bacillus Velez subtilis agent for preventing and treating Fusarium wilt as claimed in claim 1 in preparing a seedling culture medium.

5. Use of the Bacillus Velez subtilis agent for preventing and controlling Fusarium wilt as claimed in claim 1 in the preparation of biological fertilizer.

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