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Bacillus aryabhattai, microbial inoculums thereof, 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

Active Publication Date: 2017-05-31
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, reports on the degradation of beta-cypermethrin on tobacco leaves by using beta-cypermethrin degrading bacteria are rarely seen

Method used

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  • Bacillus aryabhattai, microbial inoculums thereof, preparation method and application
  • Bacillus aryabhattai, microbial inoculums thereof, preparation method and application
  • Bacillus aryabhattai, microbial inoculums thereof, preparation method and application

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preparation example Construction

[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 sus...

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 O0.02g, 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 po...

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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Abstract

The invention discloses bacillus aryabhattai, microbial inoculum thereof, a preparation method and application. The bacillus aryabhattai BC104 is beta-cypermethrin degrading bacteria, and the preservation unit is the China General Microbiological Culture Collection center (CGMCC), the preservation date is 15th, Dec, 2015, and the preservation number is CGMCC No.11664; the bacillus aryabhattai belongs to mycomonera, phylum firmicutes, bacillus class, bacilli bacteria, bacillaceae and bacillus. The microbial inoculum with the bacillus aryabhattai as an active ingredient is the microbial inoculum obtained by preforming activation culture and fermentation culture on the bacillus aryabhattai BC104. The preparation method comprises the steps of activation culture and fermentation culture; the application of the microbial inoculum is to prepare a medicine for degrading beta-cypermethrin. The strain disclosed by the invention has the advantages of simple preparation process of the microbial inoculum, low cost, convenience in use and good application prospect.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus arborii and its bacterial agent, preparation method and application. Background technique [0002] Pyrethroid insecticides belong to the third generation of pesticides, which are artificially synthesized by imitating the chemical structure of natural pyrethroids. After continuous modification and transformation of the structure of natural pyrethrins, more than 50 commercial pyrethroid pesticides have been developed, including cypermethrin, beta-cypermethrin, deltamethrin, bifenthrin, kungfuthrin, Cyfluthrin, cyhalothrin, fenpropathrin, fenvalerate, permethrin and other pyrethroid pesticides. Because pyrethroid pesticides are relatively friendly to the environment, they have a broad insecticidal spectrum, high efficacy, and high safety to mammals, so that the application of pyrethroid pesticides in health and agriculture has rapidly expanded. area. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N25/32C12R1/07
CPCA01N25/32C12N1/20C12N1/205C12R2001/07
Inventor 夏振远雷丽萍吴玉萍莫笑晗师君丽马雁军
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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