Preparation method of Bacillus strain and microbial inoculant thereof

The technology of bacillus and bacillus is applied in the preparation of bacillus and its inoculum, microbial culture, preparation of inoculum and the application field of degrading organophosphorus pesticides, which can solve the problems affecting soil quality, changing soil physical and chemical properties, and groundwater pollution. And other issues

Active Publication Date: 2015-02-25
FUZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The remaining organophosphorus pesticides will change the physical and chemical properties o

Method used

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  • Preparation method of Bacillus strain and microbial inoculant thereof
  • Preparation method of Bacillus strain and microbial inoculant thereof
  • Preparation method of Bacillus strain and microbial inoculant thereof

Examples

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example 1

[0020] Example 1: Preparation of bacillus bacterial agent

[0021] 1.1 Activation of strains

[0022] Take the Bacillus seed bacteria frozen and preserved in glycerol, streak on the LB plate, pick a single colony and inoculate it in the LB liquid medium, and place it in a shaker at 120 rpm under the conditions of aerobic, normal pressure, and 32°C. After 24 hours, expand the culture for 24 hours according to the inoculum size of 3%.

[0023] 1.2 Preparation of Bacillus bacteria agent and its effect on the degradation of organophosphorus pesticides.

[0024] The bacterium liquid of expanded culture is inserted in the fermenter by 5% inoculum amount, cultivated at 32 DEG C for 48 hours to obtain a fermented liquid, and adding mass ratio to the fermented liquid is 10% calcium carbonate and 10% sodium bicarbonate by mass ratio, Shake and shake for 5 minutes to obtain a suspension of Bacillus thallus. The solid granule bacterial agent is obtained by spray drying. The spray dryi...

example 2

[0027] Example 2: Bacillus growth and degradation conditions containing four different organophosphorus pesticide medium

[0028] Prepare 100ml of LB liquid culture medium containing acephate, methamidophos, dichlorvos, and phoxim 100mg / L respectively in 250ml Erlenmeyer flasks, and insert the activated strains into 4 bottles containing different Place in the culture medium of organophosphorus pesticides at 30°C and vibrate on a shaker at 180r / min, and compare the degradation effect after 24 hours. It was found that, in terms of cell growth, the bacillus in the four media all entered the stationary phase after 72 hours. And the cell growth condition is acephate medium>methamidophos medium>phoxim medium>dichlorvos medium.

[0029] Organophosphorus pesticide residues were detected by gas chromatography. Chromatographic conditions: inlet temperature 220°C; temperature program: stay at 120°C for 2 minutes, increase by 10°C per minute, until 250°C and keep for 30 minutes; detecto...

example 3

[0032] Example 3: Degradation of acephate by Bacillus inoculum

[0033] Insert the activated bacillus into the LB liquid medium, inoculate it into a fermenter after expanding the culture, and cultivate it at 32°C for 48 hours to obtain a fermentation broth, add calcium carbonate with a mass ratio of 10% and sodium bicarbonate with a mass ratio of 8%, The solid granule bacterial agent is obtained by spray drying.

[0034] Prepare 100ml of acephate concentration of 100mg / L solution in a 250ml Erlenmeyer flask, add the obtained bacterial agent into the Erlenmeyer flask according to the mass ratio of 1:50, place it in a shaker at 30°C and 180r / min for 24h After that, the degradation rate of acephate was measured to be 84.4%.

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Abstract

The invention provides a preparation method of a Bacillus strain and a microbial inoculant thereof. The strain is Bacillus FZUL-33 (Bacillus sp.), and was registered and collected by China General Microbiological Culture Collection Center on August 15th, 2014; and the collection number is CGMCC No.9523. The preparation method of the microbial inoculant comprises the following steps: activating and fermenting the strain to obtain a fermentation culture solution, mixing with additives, and carrying out spray drying to obtain the microbial inoculant. The microbial inoculant is used for degrading soil persistent pesticides. By using the characteristics of high temperature resistance and desiccation resistance of the Bacillus, the strain still has higher survival rate after being subjected to the microbial inoculant preparation process of spray drying; and a plate count process detects that the survival rate of the microbial inoculant is up to higher than 70% and the viable count is greater than 10<10> cfu/g.

Description

technical field [0001] The invention belongs to the field of microorganism cultivation, and in particular relates to a bacillus strain and a preparation method thereof, including microbial cultivation, preparation of the preparation and application of the degradation organophosphorus pesticide. Background technique [0002] The rapid development of contemporary agriculture inevitably leads to the emergence of pesticide residues. Among them, organophosphorus pesticides are the most widely used. Often, only a small part of the organophosphorus pesticides play an insecticidal role, while a large part remains. The remaining organophosphorus pesticides will change the physical and chemical properties of the soil, affect the soil quality, and cause groundwater pollution as the rain washes away. Organophosphorus pesticides can cause chronic poisoning to humans and animals, leading to nervous system damage. The carcinogenic, teratogenic, and mutagenic problems of pesticides are e...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10
CPCB09C1/10C12N1/20C12N1/205C12R2001/07
Inventor 程扬健林文挺韩冉许心雅黄振
Owner FUZHOU UNIV
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