Rice blast bactericide prepared by taking lysinibacillus boronitolerans as chassis cell
A technology of Bacillus lysine and rice blast fungus, which is applied in the directions of fungicides, biocides, biocides, etc., can solve the problems of excessive pesticide residues, environmental pollution, and high production costs of chemical agents, and reduce the occurrence and harm. Effect
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Embodiment 1
[0046] The separation and purification of embodiment 1 soil bacterial strain
[0047] According to an embodiment of the present application, bacteria can be isolated and purified from any soil where bio-control bacteria may exist, and their bio-control properties can be tested. According to an embodiment of the present application, tomato-rice rotation soil is taken, and the bacteria therein are separated and purified.
[0048] 1. Prepare the soil dilution. Weigh an appropriate amount of soil. The soil mentioned here is the soil that may contain biocontrol bacteria retrieved from the field in advance. According to an embodiment of the present application, the 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, break it up, and let it stand for more than 30 minutes to fully analyze the microorganisms in the soil. According to one embodiment of the presen...
Embodiment 2
[0055] Embodiment 2 identification strain type
[0056] According to an embodiment of the present application, bacterial strain species can be preliminarily identified using bacterial 16S universal primers. According to an embodiment of the present application, the bacterial strain can be directly used as a template for bacterial species identification, or the genomic DNA can be extracted first, and the genomic DNA can be used as a template 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 the agarose gel electrophoresis figure of bacterial genomic DNA extracted according to an embodiment of the application, such as figure 1As shown in A, the bands of genomic DNA are obvious, and the concentration meets the requirements of subsequent PCR experiments.
[0057] 1. Design and select primers for PCR amplification o...
Embodiment 3
[0067] The biocontrol characteristic identification of embodiment 3 isolated bacteria
[0068] According to an embodiment of the present application, the antibacterial activity of the potential biocontrol bacteria isolated in the present application can be identified by using the toxic medium method. According to one embodiment of the present application, the specific operation of the toxic medium method is:
[0069] 1. Prepare 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;
[0070] 2. Pick a single colony of boron-resistant Bacillus lysine and inoculate it in LB liquid medium, cultivate it overnight at 37°C, 150 r / min, transfer the bacterial solution the next day, and fill it with 200ml bacterial solution / 500ml LB medium amount, to a final concentration of 0.02OD 600 / ml, 150r / min rotation speed, placed in a constant temperature shaker at 37°C for about 24h, 60h and 96h, so that the OD value reached about 4....
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