A kind of bacillus arborii and its bacterial agent, preparation method and application
A technology of Bacillus arborii and Bacillus, applied in the field of microorganisms, can solve rare problems such as low cost, good application prospects, and simple preparation process
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[0030] Bacillus arzii of the present invention is the preparation method of the bacterium agent of active ingredient, comprises activation culture, fermentation culture step, specifically comprises:
[0031] A, activation culture: the Bacillus arzii ( Bacillus aryabhattai ) BC104 was inoculated into inorganic salt culture medium, and cultured by shaking for 12-24 hours at a temperature of 25-35°C and a shaking frequency of 100-250rpm to obtain liquid fermented seeds;
[0032] B. Fermentation culture: inoculate the liquid fermented seeds into the fermenter according to the volume of 1~10% of the culture medium, cultivate at 25~35°C for 24~48h, the stirring speed is 100~300rpm, the air flow rate is 0.1~1.0vvm, Get Bacillus arborii ( Bacillus aryabhattai ) BC104 fermentation broth;
[0033] C, Bacillus arborii ( Bacillus aryabhattai ) The BC104 fermentation broth was centrifuged, the supernatant was discarded, and the phosphate buffer solution with a pH value of 7.0 was suspen...
Embodiment 1
[0041] Screening and Identification of Strains
[0042] Medium:
[0043] Inorganic salt medium: NaCl 1g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH4) 2 SO 4 2.0g, MgSO 4 0.1g, 1ml trace element solution (1L trace element solution is prepared according to the following composition: MnSO 4 ·H 2 O 0.1g, ZnCl 2 0.2g, CuSO 4 ·H 2 O 0.02g, Na 2 MoO 4 2H 2 O 0.1g, made up to 1000ml with distilled water, made up to 1000ml with distilled water, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C, 30min).
[0044] Enrichment culture solution: add beta-cypermethrin solution to the inorganic salt culture solution so that the concentration of beta-cypermethrin is 50 mg / L.
[0045] LB liquid medium: Yeast powder 10g, peptone 5.0g, sodium chloride 10.0g, distilled water to make up to 1000ml, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C, 30min).
[0046] LB solid medium: Yeast p...
Embodiment 2
[0056] Bacteria preparation
[0057] 1. Inoculate the bacteria stored in the liquid test tube in the inorganic salt culture solution for activating culture for 2 days;
[0058] 2. Inoculate the activated strains in LB liquid medium, and grow them to the logarithmic growth phase at 30°C and 150rpm shaking;
[0059] 3. Centrifuge the above-mentioned bacterial solution in logarithmic growth phase for 3 minutes (6000 rpm), discard the supernatant, and suspend the bacterial cells with an appropriate amount of phosphate buffer solution with a pH value of 7.0, which is the microbial agent.
[0060] The formula of 0.2mol / L phosphate buffer solution with a pH of 7.0 is: take 39ml of 0.2mol / L sodium dihydrogen phosphate and 61ml of 0.2mol / L disodium hydrogen phosphate, dilute to 1000ml with ultrapure water, and steam under high pressure It can be obtained after sterilization (121°C, 20min).
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