Method and application of Brevibacillus laterosporus in preventing and controlling ginger blight

Abstract

The present invention relates to the field of microbial technology, and specifically to a method and application of Brevibacillus laterosporus for preventing and treating young ginger blight. The deposit number of Brevibacillus laterosporus H1 is CCTCCNO: M 20232065. The present invention discloses for the first time that Brevibacillus laterosporus H1 has the ability to prevent and treat young ginger blight and an application method thereof, providing a certain reference for later research on Brevibacillus laterosporus and having important significance for the microbial prevention and treatment of young ginger blight.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to a method and application of Brevibacillus laterosporus for preventing and treating young ginger blast. Background Art

[0002] Ginger wilt is a soil-borne disease caused by Ralstonia solanacearum. Since its discovery, domestic and foreign scholars have conducted extensive research on the infection and colonization of plants by Ralstonia solanacearum, pathogenic mechanisms, pathogenic factors, detection techniques, pathogenesis patterns, pathogenic factors and prevention and control techniques. The prevention and control methods of ginger wilt mainly fall into three categories: chemical control, agricultural control and biological control. Chemical control is the use of chemical agents to treat the cultivation soil. This method can effectively kill the pathogenic bacteria of ginger wilt, but it also causes devastating damage to the beneficial microorganisms in the soil and destroys the ecological balance of the soil. Biological control is a method of treating the cultivation soil with beneficial microorganisms or related products. This method can effectively control ginger wilt without damaging the soil. However, in the prior art, there are few reports on microbial strains for preventing and controlling ginger wilt in young ginger. Summary of the Invention

[0003] The purpose of the present invention is to provide a method and application of Brevibacillus laterosporus for preventing and treating young ginger blast.

[0004] In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:

[0005] On the one hand, the invention provides a use of Brevibacillus laterosporus in preventing and treating young ginger blast. The deposit number of Brevibacillus laterosporus H1 is CCTCCNO: M20232065.

[0006] On the other hand, a method for preventing and treating young ginger blast by Brevibacillus laterosporus is provided, comprising the following steps:

[0007] After the ginger was planted and covered with soil, the surface soil was irrigated with Brevibacillus laterosporus inoculant at a rate of 1×10 7 CFU / L, water consumption is 80L / 667m 2 -90L / 667m 2After spraying, wait for the soil surface to dry and then cover with film for heating;

[0008] 20-30 days before the young ginger is put on the market in the following year, the film is removed and the fungicide Brevibacillus laterosporus is sprayed once or twice at a concentration of 1×10 7 CFU / mL, the dosage is 2×10 12 CFU / 667m 2 -3×10 12 CFU / 667m 2 .

[0009] Furthermore, before ginger is planted, chlorine dioxide disinfectant is sprayed on the soil.

[0010] Furthermore, before digging trenches for ginger planting, chlorine dioxide disinfectant should be used for soil surface irrigation 10 days in advance. The amount of irrigation should be sufficient to moisten the surface layer of 5cm-8cm of soil. After digging trenches and before planting, chlorine dioxide disinfectant should be sprayed on the surface soil of the trench again. The water consumption is 30L / 667m 2 -35L / 667m 2 , sow seeds every 5-8 days.

[0011] Furthermore, young ginger is cultivated through greenhouse planting and heating facilities. Pipes are buried in the greenhouse and water is circulated through a boiler to increase the temperature of the ginger.

[0012] The beneficial effects of the present invention are:

[0013] The present invention discloses for the first time that Brevibacillus laterosporus H1 has the ability to prevent and treat young ginger blast and an application method thereof, which provides a certain reference for later research on Brevibacillus laterosporus and is of great significance for the microbial prevention and treatment of young ginger blast. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Schematic diagram of the colony morphology of Brevibacillus laterosporus;

[0015] Figure 2 It is the biolog identification comparison map of Brevibacillus laterosporus H1 strain;

[0016] Figure 3 This figure shows the antagonistic effect of Brevibacillus laterosporus H1 on the growth of Ralstonia solanacearum and Xanthomonas oryzae. DETAILED DESCRIPTION

[0017] The specific embodiments of the present invention are described below to facilitate understanding of the present invention by those skilled in the art. However, it should be clear that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the appended claims, these changes are obvious, and all inventions and creations utilizing the concepts of the present invention are protected.

[0018] Example

[0019] 1. Strain Source

[0020] The Brevibacillus laterosporus in this example was isolated from composted soil after the inoculation of various pathogens such as Brassica clubroot, Sclerotinia sclerotiorum, and Rhizoctonia solani. The specific isolation steps were as follows: a total of not less than 20 g of soil was collected from 5 cm below the surface of the composted soil using a five-point sampling method with a sterile key, the soil was mixed, 10 g of the soil was weighed and dissolved in 90 mL of pre-prepared sterile water, activated at 28°C, 180 rpm, and dark conditions for 30 min, and gradiently diluted to 10 8 , take 100 μL of the dilution and evenly spread it on the culture dish containing NA medium prepared in advance, culture it at 30℃ for 24h, and observe the morphology of the colonies ( Figure 1 ), pick a single colony strain for separation and purification to obtain a pure culture.

[0021] 2. Identification methods

[0022] After the strains were isolated and purified, they were compared by biolog Figure 2 ) combined with 16S rDNA sequencing, the similarity with the 16S rDNA sequence of Brevibacillus laterosporus was 99.05%, and it was identified as Brevibacillus laterosporus and named H1. The strain was deposited in the China Center for Type Culture Collection (Address: Wuhan University, Wuhan, China) on October 30, 2023, with the preservation number CCTCC NO: M 20232065.

[0023] 3. Indoor antagonistic pathogen effect

[0024] Laboratory tests have shown that Brevibacillus laterosporus H1 has a significant antagonistic effect on the growth of various pathogens. The growth inhibition rates against Botrytis cinerea, Fusarium oxysporum, and Leptosphaeria biglobosa were 39.26%, 54.46%, and 70.51%, respectively. It also had a significant inhibitory effect on the growth of Xanthomonas oryzae pv.oryzae and Ralstonia solanacearum, and was significantly better than strains A1, TN4, and B22. Figure 3 The transparent zone (antagonistic zone) formed by inhibiting the growth of Ralstonia solanacearum, the pathogen of ginger blast, reached 8 mm, and the growth inhibition effect was significant.

[0025] Test example

[0026] This test case is to plant young ginger in a continuous cropping mode and off-season. Specifically, young ginger is cultivated through greenhouse planting and heating facilities. Pipes are buried in the greenhouse, and water is circulated through the boiler to heat the ginger and maintain the ground temperature at around 25°C. This planting method allows ginger to be sown in October and the young ginger can be harvested after New Year's Day. The wholesale price is more than 5 times higher than that of ginger in the later period. However, the cultivation of young ginger in heating facilities is different from field cultivation. It is inconvenient to apply pesticides. During the heating period, the insulation film cannot be removed for pesticide application. Pesticides can only be applied before planting and after the heating is stopped. If chemical control is applied, the application of pesticides before planting is ineffective. If the pesticides are applied 20 days before listing, the time is short and the effect cannot be shown. In addition, there is a hidden danger of pesticide residues. In order to cooperate with the cultivation of young ginger in heating facilities, to achieve the prevention and control of young ginger plague and increase the yield of young ginger, the method of preventing and controlling young ginger plague in this test case is as follows:

[0027] Ten days before digging trenches for ginger planting, treat the soil surface with a 1000-fold dilution of chlorine dioxide disinfectant (suitable for livestock enclosures). The ideal amount of water should be sufficient to moisten the top 5 cm of soil. After digging trenches and before planting, spray the surface soil with chlorine dioxide disinfectant again, using 30 L / mu. Leave 5-8 days between plantings.

[0028] After the ginger was planted and covered with soil, the surface soil was irrigated with Brevibacillus laterosporus H1 to further inhibit the growth of ginger wilt pathogens and regulate the soil microbial community structure. The application rate was 1×10 7 CFU / L, water consumption is 90L / 667m 2After spraying, wait for the soil surface to dry and then cover it with film for heating.

[0029] 20-30 days before the young ginger is put on the market in the following year, the film is removed and the Brevibacillus laterosporus H1 fungicide is sprayed once or twice at a concentration of 1×10 7 CFU / mL, the dosage is 2×10 12 CFU / 667m 2 .

[0030] At the same time, only chlorine dioxide disinfectant, only Brevibacillus laterosporus H1, conventional chemical control, and CK (no control) were set as control groups. The effects of preventing and controlling young ginger blight and increasing young ginger yield are shown in Table 1 below.

[0031] Table 1 Effects of different treatments on preventing and controlling ginger blight and increasing ginger yield

[0032]

[0033] Note: The average original planting amount of ginger is about 20t / mu, or 30kg / m 2 .

[0034] As can be seen from the table above, Brevibacillus laterosporus H1 is effective in preventing and controlling ginger blight and increasing the yield of early-growing ginger. This study first discloses that Brevibacillus laterosporus has the ability to prevent and control ginger blight in young ginger, providing a reference for future research on Brevibacillus laterosporus and having important implications for the microbial control of ginger blight in young ginger.

[0035] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the present invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

Claims

1. The application of Brevibacillus laterosporus in preventing and treating ginger blight of young ginger is characterized in that: The deposit number of Brevibacillus laterosporus H1 is CCTCC NO: M20232065.

2. The method for preventing and treating young ginger blast with Brevibacillus laterosporus H1 according to claim 1, characterized in that: The following steps are involved: After the ginger was planted and covered with soil, the surface soil was irrigated with Brevibacillus laterosporus H1 inoculant at a dosage of 1×10 7 CFU / L, water consumption is 80L / 667m 2 -90L / 667m 2 After spraying, wait for the soil surface to dry and then cover with film for heating; 20-30 days before the young ginger is put on the market in the following year, the film is removed and the Brevibacillus laterosporus H1 fungicide is sprayed once or twice at a concentration of 1×10 7 CFU / mL, the dosage is 2×10 12 CFU / 667m 2 -3×10 12 CFU / 667m 2 .

3. The method for preventing and treating young ginger blast with Brevibacillus laterosporus H1 according to claim 2, characterized in that: Before planting ginger, spray chlorine dioxide disinfectant on the soil.

4. The method for preventing and treating young ginger blast with Brevibacillus laterosporus H1 according to claim 3, characterized in that: Before digging trenches for ginger planting, 10 days in advance, irrigate the soil surface with chlorine dioxide disinfectant. The amount of irrigation should be enough to moisten the surface 5cm-8cm of the soil. After digging trenches and before planting, spray the surface soil in the trench with chlorine dioxide disinfectant again. The water consumption is 30L / 667m 2 -35L / 667m 2 , sow seeds every 5-8 days.

5. The method for preventing and treating young ginger blast with Brevibacillus laterosporus H1 according to claim 2, characterized in that: Young ginger is cultivated through greenhouse planting and heating facilities. Pipes are buried in the greenhouse and water is circulated through a boiler to increase the temperature of the ginger.

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