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Biocontrol strain BS71 and application thereof to preventing crops from being polluted by aflatoxin

A technology of aflatoxin and bio-control strains, which is applied in the field of crop disease prevention and control, can solve problems such as poor effects, achieve good control effects, improve product quality, and achieve good control effects

Active Publication Date: 2014-05-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The Chinese patent literature with publication number CN102816725A also discloses a kind of Bacillus subtilis T-500, although this bacterial strain has antagonistic effect to various pathogenic bacteria, the inhibition rate to Aspergillus flavus is only 35.7%, and the effect is relatively poor.

Method used

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  • Biocontrol strain BS71 and application thereof to preventing crops from being polluted by aflatoxin
  • Biocontrol strain BS71 and application thereof to preventing crops from being polluted by aflatoxin
  • Biocontrol strain BS71 and application thereof to preventing crops from being polluted by aflatoxin

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Experimental program
Comparison scheme
Effect test

Embodiment 1B

[0035] The collection, separation and identification of embodiment 1BS71 biocontrol strain

[0036] BS71 strain was isolated from peanut rhizosphere soil in Shaoxing City, Zhejiang Province. Take the soil and put it in sterile water, shake it for 1 hour, absorb the supernatant, put it in a water bath at 80°C for 10 minutes, and spread it on an LB plate; pick colonies for purification and culture the next day.

[0037] The colony surface of strain BS71 on LB medium is rough, opaque, shrunken and white (see figure 1 ), able to form spores, mobile, aerobic.

[0038]The DNA of the strain was extracted, the 16S rDNA sequence was amplified, and sequenced. The 16S rDNA of the strain was shown as SEQ ID No.1, and the sequence comparison results in NCBI showed homology with Bacillus subtilis Highest.

[0039] The strain has been preserved in the General Microbiology Center (CGMCC) of the China Committee for Microbial Culture Collection (CGMCC), located at No. 3, No. 1, Beichen West ...

Embodiment 2

[0040] The preparation of embodiment 2 biocontrol agents

[0041] 1. Preparation of biocontrol agents

[0042] (1) Inoculate the biocontrol strain BS71 into LB culture medium, culture at 30°C with 180r / min shaking, take samples in the ultra-clean workbench after 12 to 16 hours and measure the OD value at 600nm. Bacteria culture solution to zero;

[0043] (2) When the OD value of the bacterial liquid is between 0.5 and 0.8, it is the seed bacterial liquid, and the seed bacterial liquid is added to a 200L fermenter with a fermentation medium at a volume ratio of 1:100, at 30°C 250r / min shaking culture for 20-24 hours, the pH value of the fermentation broth is maintained at 7.0, the dissolved oxygen is 20%, and the ventilation volume is 5-7m 3 / hour, the tank pressure is 0.05-0.1KPa. After the fermentation broth is out of the tank, the gradient dilution plate is used to detect and adjust the concentration of the bacterial suspension to about 1×10 9 cfu / ml.

[0044] The form...

Embodiment 3B

[0045] Antagonistic activity determination of embodiment 3BS71 biocontrol strains to Aspergillus flavus

[0046] 1. Determination of antagonistic activity of BS71 against Aspergillus flavus plate

[0047] (1) Test method

[0048] Using the confrontation culture method, inoculate the Aspergillus flavus HZ17 (a strain producing aflatoxin identified in the previous test, which is resistant to pyrimethanil) stored at 4°C on a PDA plate for activation, and wait until the fungus is overgrown on the plate. Use a sterilized puncher to evenly punch out circular bacterial blocks with a diameter of 6 mm from the outer edge of the colony. Inoculate the mycelium block on the center of the WA plate (WA medium / L: peptone 5g, glucose 10g, gravy extract 3g, sodium chloride 5g, agar 20g, pH=7.0), and place them around 25mm away from the center respectively. BS71 was inoculated, and the test took Bacillus subtilis type strain PY79, clear water and 10 μg / ml pyrimethanil as the control group. C...

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Abstract

The invention discloses a biocontrol strain BS71 and an application thereof to preventing crops from being polluted by aflatoxin. The biocontrol strain BS71 is bacillus subtilis BS71, with collection number of CGMCC No.8191. The application method comprises the steps of preparing a biocontrol agent of the biocontrol strain BS71 and applying the biocontrol agent to the crops. The biocontrol strain has strong antagonistic action on aspergillus flavus and can be used for preventing pollution caused by aflatoxin. The invention provides a biocontrol method for pollution caused by aflatoxin. As chemical pesticides are not used, the biocontrol strain has the advantages of no toxicity, low possibility of having resistance to drugs, environmental and biological safety and the like, and has extensive application prospects.

Description

technical field [0001] The invention belongs to the technical field of crop disease prevention and control, and in particular relates to a biocontrol strain BS71 and its application in preventing and controlling aflatoxin pollution of crops. Background technique [0002] Aflatoxin (hereinafter abbreviated as AFT) is mainly a metabolite of Aspergillus flavus and A. parasiticus, and is the most toxic type of biotoxin found so far in contaminated agricultural products. According to the World Food and Agriculture Organization estimates, currently 25% of the world's crop products are contaminated by mycotoxins, mainly AFT. In my country, the aflatoxin pollution of peanuts, corn and other crops has seriously affected food safety and the export competitiveness of agricultural products. [0003] Due to the huge economic losses and health hazards caused by AFT pollution, people have been exploring effective measures to control AFT pollution. Breeding pest-resistant varieties is one...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/125
Inventor 陈云尹燕妮马忠华
Owner ZHEJIANG UNIV
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