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Antibiological inoculant for toxin production aspergillus flavus, and preparation method and application of antibiological inoculant

A technology of Aspergillus flavus and biocontrol agent, which is applied in biochemical equipment and methods, methods of using spores, and botanical equipment and methods, etc. Field growth, poor adaptability of a single bacterial agent, etc., to achieve the effect of less aflatoxin content, significant pollution effect, and significant control effect

Active Publication Date: 2019-08-02
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are some bio-control bacteria that are not suitable for field growth, have no growth advantage in the field, and cannot achieve the effect of inhibiting toxin-producing Aspergillus flavus
Moreover, a single bacterial agent has the problems of poor adaptability and poor control effect.

Method used

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  • Antibiological inoculant for toxin production aspergillus flavus, and preparation method and application of antibiological inoculant
  • Antibiological inoculant for toxin production aspergillus flavus, and preparation method and application of antibiological inoculant
  • Antibiological inoculant for toxin production aspergillus flavus, and preparation method and application of antibiological inoculant

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Experimental program
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Effect test

Embodiment 1

[0034] The acquisition of embodiment 1 bacterial strain PEAS-10 and PAF-1

[0035] 1. Isolation, purification and identification of strains

[0036] 1. Sample collection: Samples were collected at the peanut planting site (the sampling time was 2018.05, and the sampling sites were Tangcunba Village, Tumen Township, Jialing District, Nanchong City, Sichuan Province, and Wangcheng Sub-district Office, Laixi City, Qingdao City, Shandong Province). Sample (100g) Take 5 sub-samples (2cm wide, 5cm deep soil) in the range of 10×10m according to the diagonal method, and mix them as one sample. Put the collected samples into plastic bags, prick some pinholes to facilitate gas exchange, transport them to the laboratory, and store them at 4°C for screening of Aspergillus flavus.

[0037] 2. Isolation and purification of bacterial strains.

[0038] (1) Preparation of soil sample bacterial suspension

[0039] Take 10g soil sample, add 90mL 0.1% peptone sterile water (w / v), shake at room...

Embodiment 2

[0072] Example 2 Analysis of the toxin production of Aspergillus flavus PEAS-10 and PAF-1

[0073] (1) Toxigenic culture

[0074] Aspergillus flavus PEAS-10 and PAF-1 strains were respectively inoculated on the MEA slant test tube culture medium, and cultured at 28°C for 3 days to activate them; 4mL sterile water was added to the slant test tube culture medium and rinsed to prepare yellow koji Mold PEAS-10 suspension and Aspergillus flavus PAF-1 suspension. Record the number of spores under a microscope with a hemocytometer.

[0075] Add 10mL of toxin-producing culture solution to a 50mL centrifuge tube, and then add a certain amount of Aspergillus flavus PEAS-10 or PAF-1 bacterial suspension, so that the final concentration of spores is 10 5 / mL, 30°C, 200rpm, cultivated for 7 days.

[0076] (2) Aflatoxin B in the toxin-producing culture medium 1 Determination of

[0077] The methods of immunoaffinity chromatography purification, liquid chromatography separation, and flu...

Embodiment 3

[0081] A kind of biocontrol agent for preventing and controlling aflatoxin pollution and preparation method thereof

[0082] (1) Microorganisms used: Aspergillus flavus PEAS-10 and Aspergillus flavus PAF-1

[0083] (2) Strain activation: the strains were respectively inoculated on MEA medium, and cultured at 30° C. for 3-5 days until yellow-green spores were produced.

[0084] (3) Preparation of bacterial culture medium: crush peanut red coat to a size of about 0.5×0.5 cm, mix peanut red coat and distilled water at a ratio of 1:1-2:3, and add 1% by mass at the same time -1.5% CaCl 2 , sterilized at 121°C for 20 minutes.

[0085] (3) Inoculate the activated non-toxin-producing Aspergillus flavus strains onto the sterilized inoculum medium respectively, cultivate them for 5-8 days at 30° C., and shake once a day to make Aspergillus flavus grow evenly on the medium; After 5-8 days of cultivation, the number of spores of Aspergillus flavus reached 10 8 / g medium or more.

[0...

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Abstract

The invention discloses an antibiological inoculant for toxin production aspergillus flavus, and a preparation method and an application of the antibiological inoculant, and belongs to the technical field of the antibiological inoculant for harmful microorganisms. The effective ingredients of the antibiological inoculant for the toxin production aspergillus flavus are aspergillus flavus PEAS-10 producing no aflatoxin and aspergillus flavus PEF-1 producing no aflatoxin; a aspergillus flavus strain in the antibiological inoculant can rapidly grow and propagate in the field, the growth, the propagation and the toxin production of the toxin production aspergillus flavus are efficiently inhibited, the disease of a peanut is obviously reduced, the content of the aflatoxin in the harvested peanutis low, the storage period of the peanut is long, and compared with the treatment with a single microbial agent, the effect is excellent.

Description

technical field [0001] The invention belongs to the technical field of biocontrol agents for harmful microorganisms, and in particular relates to a biocontrol agent for toxin-producing Aspergillus flavus, a preparation method and application thereof. Background technique [0002] Aflatoxins are metabolites of Aspergillus flavus and Aspergillus parasiticus. A large number of experimental data show that aflatoxin can induce experimental liver cancer in humans and various animals. It is the strongest chemical carcinogen discovered so far, and its ability to induce liver cancer is 75 times greater than that of dimethylnitrosamine. Aflatoxin is also a highly toxic substance. Its acute toxicity is 68 times that of arsenic and 10 times that of potassium cyanide. It can seriously damage the liver and cause death in a short period of time. [0003] Peanuts are the most susceptible crops to be infected with Aspergillus flavus. Peanuts are likely to be infected with Aspergillus flavu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/04A01P3/00A01P21/00C12N1/14C12N3/00C12R1/67
CPCC12N1/14C12N3/00A01N63/30
Inventor 孙杰于丽娜张建成张初署王明清毕洁杨伟强栾云霞
Owner SHANDONG PEANUT RES INST
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