Areca nut root rot fungicide taking lysinibacillus boronitolerans as base cells
A technology of Bacillus lysinus and root rot bacteria, applied in the direction of fungicides, biocides, biocides, etc., can solve the problems of excessive pesticide residues, environmental pollution, and high production costs of chemical agents, and achieve the effect of maintaining production
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Embodiment 1
[0044] Example 1, isolation and purification of soil strains
[0045] According to an embodiment of the present application, bacteria can be isolated and purified from any soil where biocontrol bacteria may exist, and their biocontrol properties can be tested. According to an embodiment of the present application, tomato-rice rotation soil is taken, and the bacteria in it are isolated and purified.
[0046] 1. Prepare a soil dilution. Weigh an appropriate amount of soil. The soil mentioned here is the soil that may contain biocontrol bacteria that was retrieved from the wild in advance. According to an embodiment of the present application, soil can also be directly weighed in the field. According to an embodiment of the present application, 10 g of soil can be weighed. Put the weighed soil into sterile water, crush it, and let it stand for more than 30 minutes to fully analyze the microorganisms in the soil. According to an embodiment of the present application, 10 g of ...
Embodiment 2
[0053]Example 2, identification of strain species
[0054] According to an embodiment of the present application, the strain species can be preliminarily identified by using the 16S universal primer. According to an embodiment of the present application, strains can be directly used as substrates for bacterial species identification, or genomic DNA can be extracted first, and genomic DNAs can be used as substrates for bacterial species identification. According to an embodiment of the present application, agarose gel electrophoresis is used to detect the content and quality of the extracted bacterial genomic DNA. figure 1 A is an agarose gel electrophoresis image of bacterial genomic DNA extracted according to an embodiment of the present application, such as figure 1 As shown in A, the bands of genomic DNA are obvious, and the concentration meets the requirements of subsequent PCR experiments.
[0055] 1. Design and select primers for PCR amplification of 16S rDNA. Accordi...
Embodiment 3
[0064] Example 3, identification of biocontrol properties of purified bacteria
[0065] According to an embodiment of the present application, the antibacterial activity of the potential biocontrol bacteria isolated by the present application can be identified by using a toxic medium method. According to an embodiment of the present application, the specific operation of the poisonous medium method is:
[0066] 1. Configure PDA solid medium and cool it to about 55°C; or heat and melt the solidified PDA medium and cool it to about 55°C;
[0067] 2. Pick a single colony of borolysine-resistant Bacillus and inoculate it in LB liquid medium, cultivate overnight at 37°C, 150 r / min, transfer the bacterial liquid the next day, and use 200 ml bacterial liquid / 500 ml LB medium for filling amount to a bacterial concentration of 0.02OD 600 / ml, 150r / min speed, placed on a constant temperature shaker at 37°C for about 24h, 48h, 72h, 96h, 120h, so that the OD 600 The fermentation produc...
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