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Aspergillus flavus strain without producing aspergillus flavus toxin and uses thereof

A technology for aflatoxins and strains of Aspergillus flavus, which can be used in applications, fungicides, fungi, etc., can solve the problem of not having a good inhibitory effect, and achieve the effect of reducing the risk of aflatoxin contamination and high competitiveness.

Inactive Publication Date: 2009-02-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biocontrol strains in the United States do not have a good inhibitory effect on the African A. flavus population, while the highly competitive non-toxin-producing strains isolated from Africa have a good inhibitory effect on the African toxigenic A. flavus; it shows that different The non-toxin-producing Aspergillus flavus biocontrol strains in the region have a certain scope of application
At present, there are no relevant reports in this field in our country

Method used

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  • Aspergillus flavus strain without producing aspergillus flavus toxin and uses thereof
  • Aspergillus flavus strain without producing aspergillus flavus toxin and uses thereof
  • Aspergillus flavus strain without producing aspergillus flavus toxin and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, collection, separation, identification of biocontrol bacterial strain A051 (being Aspergillus flavus strain A051)

[0027] (1), isolate Aspergillus flavus strain from soil

[0028] Aspergillus flavus strains were isolated from the soil collected from peanut fields in Jiangsu, Zhejiang, Shandong, etc. with YES medium. Per liter of YES medium composition: Yeast extract 1g, sucrose 10g, NaCl 60g, agar 20g, add CuSO4.ZnSO41ml, 0.4% nitramine 5ml, streptomycin 100ppm after sterilization.

[0029] (2), use bioassay, thin-layer chromatography and molecular biological method to screen out the aspergillus flavus strain that does not produce AFT wherein, measure the growth rate of these not producing AFT bacterial strains, sporulation amount, the survival period in soil then and Competitiveness with AFT-producing strains and other indicators, thus screening a highly competitive non-AFT-producing Aspergillus flavus strain A051. details as follows:

[0030]Bioassay...

Embodiment 2

[0034] Example 2, Deletion Determination of Strain A051 Toxin Synthesis Gene

[0035] (1) Primer synthesis

[0036] The present invention is based on the sequence of the toxin synthesis gene of Aspergillus flavus with Genbank accession number AY510451, and designs the detection of toxin synthesis related genes C1, C3, norB, cypA, aflT, pksA, aflR, norA, ver1, verA, ordA, hypA And glcA primers, primer sequences and amplified fragment size are shown in Table 1.

[0037] Table 1. Primer sequences of different toxin synthesis genes

[0038] Gene

Primer name

Sequence (5'-3')

The amplified fragment is large

small C1

C1-F

C1-R CCGTTGCAGTTAAAACTCACA (SEQ ID NO: 1)

GAAGCTCGATCTTCTCCATGA (SEQ ID NO: 2) 409

C3

C3-F

C3-R TCTGGAGTCGGAGGTTAGGTT (SEQ ID NO: 3)

GAGCAACACGATCATTGCAT (SEQ ID NO: 4) 544

norB

cypA norB-cypA-F

norb-cypA-R GTGCCCAGCATCTTGGTCCA (SEQ ID NO: 5)

AGGACTTGATGATTCCTCGTC (SEQ ID NO: 6) ...

Embodiment 3

[0044] Example 3, Determination of the Deletion Site Sequence of the Toxin Synthesis Gene of Bacterial Strain A051

[0045] It has been determined by the above method that the strain A051 is deleted from the first toxin synthesis gene to the toxin synthesis gene norA, and is complete from the ver1 gene to the last toxin synthesis gene. According to the sequence of ver1, three specific nested primers AFT1, AFT2 and AFT3 were designed and combined with one random primer RA5 for Tail-PCR amplification to determine the specific deletion site. (See Table 2 for primer sequences)

[0046] Table 2. Sequences of nested primers and random primers used in Tail-PCR amplification

[0047] Primer name Primer sequence (5'-3') AFT1 CATCGGCCTGGATTGCGATA (SEQ ID NO: 27) AFT2 ACTTTCTCCGCGGCCTCAC (SEQ ID NO: 28) AFT3 CGCCGGTGACTAAGGCCACT (SEQ ID NO: 29) RA5 WGTGNAGWANCANAGA

[0048] Carry out nested PCR reactions according to the following steps, each...

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PUM

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Abstract

The invention discloses an Aspergillus flavus strain for not producing toxins of Aspergillus flavus, the Aspergillus flavus strain belongs to the Aspergillus flavus of Aspergillus, the collection name is: Aspergillus flavus strain A051, the collection unit is: China General Microbiological Culture Collection Center, the collection data is: June 24, 2008, and the collection No. is: CGMCC NO.2556. The invention further discloses the use of the Aspergillus flavus strain, and the use is characterized in that: the Aspergillus flavus strain is used for inhibiting Aspergillus flavus pathogenic bacteria groups of the toxins of the Aspergillus flavus in peanut field soil.

Description

technical field [0001] The invention belongs to the field of agricultural product safety, and mainly relates to a biocontrol bacterial strain, Aspergillus flavus strain A051, which inhibits aflatoxin-producing Aspergillus flavus populations in peanut field soil, and the biocontrol application of the bacterial strain. Background technique [0002] Aflatoxins (hereinafter abbreviated as AFT) are mainly metabolites of Aspergillus flavus and A. parasiticus, and are the most toxic type of biotoxins found so far in contaminated agricultural products. Among them, aflatoxin B1 is the most toxic and harmful, and its toxicity is 10 times that of potassium cyanide and 68 times that of arsenic. It is listed as a highly toxic substance under strict control. AFT pollution of agricultural products has become an invisible killer affecting food safety and endangering people's health. [0003] According to the World Food and Agriculture Organization estimates, currently 25% of the world's cr...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/67
Inventor 马忠华蒋金花严蕾艳尹燕妮
Owner ZHEJIANG UNIV
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