Aspergillus flavus producing no aflatoxin and application thereof

A technology of aflatoxin and aspergillus flavus, which is applied in the directions of application, fungi, method of using spores, etc., to achieve the effect of reducing aflatoxin pollution

Active Publication Date: 2014-01-15
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, non-toxin-producing strains from different regions have a certain scope of application in inhibiting toxin-producing bacteria. For example, the non-toxin-producing bacteria screened in Africa can better inh...

Method used

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  • Aspergillus flavus producing no aflatoxin and application thereof
  • Aspergillus flavus producing no aflatoxin and application thereof
  • Aspergillus flavus producing no aflatoxin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, collection, separation, identification of Aspergillus flavus strain GZ-17

[0045] 1. Isolation of Aspergillus flavus strain GZ-17 from peanut planting soil

[0046] Isolation of Aspergillus flavus strains from Guangdong peanut planting soil with DG18 medium. The specific operation is as follows:

[0047] 1. Preparation of soil sample bacterial suspension

[0048] Take 10g of soil sample, add 90mL of 0.1% peptone sterile water (w / v), shake at room temperature for 30min, and make 10 -1 bacteria suspension; then take 0.5mL10 -1 Add 4.5mL0.1% peptone sterile water to the bacterial suspension to prepare 10 -2 Bacteria suspension of dilution; prepare 10 as above -3 diluted bacterial suspension.

[0049] 2. Isolation and purification of strains

[0050] Take 0.1mL bacterial solution for each dilution, spread on DG18 medium (recipe: 5.0g casein peptone, 10.0g anhydrous glucose, 1.0g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.002g clonamine, ...

Embodiment 2

[0068] Example 2. Deletion identification of aflatoxin synthesis gene in Aspergillus flavus (Aspergillus flavus) GZ-17CGMCC No.8050

[0069] The inventors of the present invention took the sequence of the toxin synthesis gene of Aspergillus flavus with the Genbank accession number AY510451 as a reference, and designed four pairs of primers for the toxin synthesis related genes hypB, hypC, hypD and hypE. The primers of other genes refer to Perng-Kuang Chang (Chang , P.K., Horn, B.W. and Dorner, J.W. (2005) Sequence breakpoints in the aflatoxin biosynthesis gene cluster and flanking regions in nonaflatoxigenic Aspergillus flavus isolates. Fungal Genet Bio42, 914–923.). The 29 genes to be tested and their corresponding primer sequences are shown in Table 1.

[0070] Table 1 Primer sequences of different aflatoxin synthesis genes

[0071]

[0072]

[0073]

[0074] Extract the genomic DNA of Aspergillus flavus (Aspergillus flavus) GZ-17CGMCC No.8050, and use it as a templa...

Embodiment 3

[0079] Example 3. Inhibition of Aspergillus flavus (Aspergillus flavus) GZ-17CGMCC No.8050 on PDA medium to produce aflatoxin-producing Aspergillus flavus

[0080] 1. Experimental method

[0081] Prepare spores at a concentration of 10 4 、10 5 、10 6 pcs / ml Aspergillus flavus (Aspergillus flavus) GZ-17CGMCC No.8050 and 10 5 aflatoxin-producing Aspergillus flavus GD-1 spore suspension, the two were mixed at a volume ratio of 1:1, and after mixing, 30 μl of the spore suspension was added to a 6cm PDA medium (formula: Potato powder 6.0g / L, glucose 20.0g / L, agar 20.0g / L) plate, cultivated at 30°C for 10 days, observed the colony morphology, and determined the content of aflatoxin in it by ultra-high performance liquid chromatography.

[0082] Among them, the aflatoxin (in AFB) was determined by ultra-high performance liquid chromatography 1 As an example) the content method is as follows:

[0083] (1) Preparation of samples to be tested

[0084] After culturing at 30°C for 1...

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Abstract

The invention discloses an aspergillus flavus strain producing no aflatoxin. The aspergillus flavus strain producing no aflatoxin, provided by the invention, is specifically aspergillus flavus GZ-17; the preservation number of the aspergillus flavus strain in the general microbiology center of the China Committee for Culture Collection of Microorganisms is CGMCC No.8050. Experimental results show that the aspergillus flavus strain GZ-17 provided by the invention plays a role in restraining the poison production of the aspergillus flavus strain producing the aflatoxin; when the concentration ratio of the strain to a toxic producing fungus spore is 105: 105, the poison production restrained ratio of bacterium producing no poison to the bacterium producing poison is nearly 96%. The strain has important significance for restraining the infection of the aspergillus flavus producing poison to agricultural products and reducing the aflatoxin pollution in the agricultural products.

Description

technical field [0001] The invention belongs to the field of agricultural product safety, and relates to a strain of Aspergillus flavus that does not produce aflatoxin and the application of the strain in inhibiting toxin production by Aspergillus flavus. Background technique [0002] Aflatoxin (AFT), a class of secondary metabolites mainly produced by fungi such as Aspergillus flavus and Aspergillus parasiticus, has been isolated from AFB 1 、AFB 2 、AFG 1 、AFG 2 、AFM 1 、AFM 2 18 kinds of aflatoxins with different structures, the most important of which is AFB 1 、AFB 2 、AFG 1 、AFG 2 . In my country, aflatoxin is mainly produced by Aspergillus flavus. Aflatoxins have carcinogenic, teratogenic, and mutagenic effects, and are identified as a first-class carcinogen by the World Health Organization (WHO), which seriously affects human health. Among them, AFB 1 Potassium is the most toxic, and its toxicity is 10 times that of potassium cyanide and 68 times that of arseni...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N3/00A23L1/015C12R1/67A23L5/20
Inventor 刘阳魏丹丹周露张初署邢福国赵月菊王龑
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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