Bacillus tequilensis and application thereof
A technology of Bacillus tequila and Bacillus, applied in the application, bacteria, fungicides and other directions, can solve the problems of low colonization and proliferation ability, narrow antibacterial spectrum, low control effect, etc. Simple preparation method and low cost effect
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Embodiment 1
[0071] The acquisition and identification of embodiment 1 Tequila bacillus 2-2a
[0072] 1. Screening and isolation of Bacillus tequila 2_2a strain
[0073] (1) Sample collection: collect fresh leaves of soybean plants in the base of Cangzhou Academy of Agricultural Sciences, Hebei Province, rinse the surface with dust with sterile water, and then carry out surface disinfection of plant leaves (soak with 75% alcohol for 1 min and 8% Soak in NaClO for 4min to carry out surface disinfection treatment), wash with aseptic water 4 times;
[0074] (2) Separation and screening: cut the leaves into 1cm×1cm pieces, add water and grind them to a paste, let them stand for 10 minutes, spread them on LB plates, and culture them at 30°C for 48 hours;
[0075] (3) Purification: After the culture grows out, use the plate streaking separation method to purify, pick the colonies that grow at the highest concentration and streak on the enrichment medium until the pure culture is isolated.
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Embodiment 2
[0096] The preparation of embodiment 2 bacillus tequila 2-2a bacterium liquid and bacterial agent
[0097] (1) Strain activation: Pick the Bacillus tequila 2-2a bacterial strain to the strain preservation medium, after continuous streaking at 30°C and picking a single colony for two times, pick a single colony in the strain activation medium, and 30°C, 180r / min shaker shaking culture for 24h;
[0098] (2) liquid seed preparation: in the fermenter that the seed culture medium of high temperature sterilization is housed, according to the 2-2a activation bacterial liquid that the inoculum size of 10% (V / V) obtains inoculation step (1), at 30 ℃, pass Air culture for 18 hours to obtain liquid seeds;
[0099] (3) Liquid state fermentation: In the seed culture medium equipped with high-temperature sterilization, inoculate liquid seeds according to the inoculum size of 10% by volume fraction, at 30 ℃, cultivate 24h (logarithmic growth phase) under 180r / min, obtain viable bacteria So...
Embodiment 3
[0101] The bacteriostasis of embodiment 3 Tequila bacillus 2-2a
[0102] The present embodiment analyzes Tequila bacillus 2-2a to soybean anthracnose bacterium (Colletotrichum chlorophyti), mouth protruding umbilicus (Exserohilum rostratum), Alternaria alternata (Alternaria alternata), Phoma herbarum (Phoma herbarum), gray spot fungus (Cercospora sojina), Fusarium oxysporum (Fusarium oxysporum) and Fusarium equiseti (Fusarium equiseti), the specific methods are as follows:
[0103] (1) Soybean anthracnose fungus (Colletotrichum chlorophyti), mouth protruding umbilical cordworm (Exserohilum rostratum), Alternaria alternata (Alternaria alternata), Phomaherbarum (Phomaherbarum), gray spot fungus (Cercospora sojina), Fusarium oxysporum Fungus (Fusarium oxysporum) and Fusarium equiseti (Fusarium equiseti) pathogens were inoculated on PDA plates respectively, and after being cultured at 25°C to cover the plates, bacteria cakes were made with a hole punch with a diameter of 5 mm, and...
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