Antagonistic agent for toxigenic Aspergillus flavus, preparation method and application thereof
A technology of aspergillus flavus and antagonistic antimicrobial agent, which is applied in the field of antagonistic antimicrobial agents of harmful microorganisms, can solve the problems of inability to inhibit toxin-producing aspergillus flavus, poor adaptability of a single agent, and unsuitability for field growth, etc., and achieve aflatoxin The effect of less content, significant control effect, and increased yield
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Embodiment 1
[0034] The acquisition of embodiment 1 bacterial strain PEAS-10 and PEASH-12
[0035] 1. Isolation, purification and identification of strains
[0036] 1. Sample collection: Samples were collected at the peanut planting site (2018.05, the sampling sites were Tangcunba Village, Tumen Township, Jialing District, Nanchong City, Sichuan Province, and Hongqi Branch of Jiangxi Provincial Fine Breeding Farm). Each sample (100g) was collected at 10 Take 5 sub-samples (soil with a width of 2 cm and a depth of 5 cm) according to the diagonal method in the range of ×10m, 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 ro...
Embodiment 2
[0072] Example 2 Analysis of the toxin production of Aspergillus flavus PEAS-10 and PEASH-12
[0073] (1) Toxigenic culture
[0074] Aspergillus flavus strains PEAS-10 and PEASH-12 were inoculated on the MEA slant test tube culture medium respectively, 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 PEASH-12 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 PEASH-12 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 separat...
Embodiment 3
[0081] Antagonist for preventing and controlling aflatoxin pollution and preparation method thereof
[0082] (1) Microorganisms used: Aspergillus flavus PEAS-10 and Aspergillus flavus PEASH-12
[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 shells to a size of about 0.5×0.5 cm, mix them according to the mass ratio of peanut shells and distilled water in a ratio of 1:1-2:3, and add 1%-1.5 %CaCl 2 and 0.5% KCl, sterilized at 121°C for 20min.
[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 more than one / g culture ...
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