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Enterobacter cloacae biocontrol strain for efficiently inhibiting aspergillus flavus compounded aflatoxin and its application

A technology of Enterobacter cloacae and aflatoxin, which is applied in the field of microorganisms to achieve the effect of preventing and controlling aflatoxin pollution

Active Publication Date: 2018-03-23
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current existing research, there is no report on the inhibition of Aspergillus flavus by non-decarboxylated Lerekia

Method used

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  • Enterobacter cloacae biocontrol strain for efficiently inhibiting aspergillus flavus compounded aflatoxin and its application
  • Enterobacter cloacae biocontrol strain for efficiently inhibiting aspergillus flavus compounded aflatoxin and its application
  • Enterobacter cloacae biocontrol strain for efficiently inhibiting aspergillus flavus compounded aflatoxin and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1) Activate Enterobacter cloacae 3J1EC on the LB plate, culture it in a 37°C incubator for 24 hours, pick up a single colony of the activated Enterobacter cloacae with a pick, and transfer it to a Erlenmeyer flask containing 15 mL of LB liquid medium , Shaking culture at 28°C, 200r·min-1 for 12h. Aspirate 1% of the culture solution and transfer it to a Erlenmeyer flask filled with 15mL LB liquid medium for culture, and culture with shaking at 28°C and 200r·min-1 for 12h to obtain the fermentation solution of the antagonistic strain.

[0019] 2) Enterobacter cloacae fermentation broth (final concentration 1×10 7 CFU / mL) and the vigorously growing Aspergillus flavus suspension (the final concentration of spores was 5.0×10 5 Spores mL-1) were cultured together in Sabouraud liquid medium at 28°C and 200 rpm for 5 days, and each treatment was repeated 3 times.

[0020] 3) Measure the content of aflatoxin B1 in its culture solution (Table 2).

[0021] Table 2 Control effec...

Embodiment 2

[0025] 1) Take Zhonghua No. 6 peanut grains from the peanut field in Hubei and grind them into powder, weigh 1g of peanut powder in a petri dish, and add 1ml of Aspergillus flavus spore liquid (5×10 5 cells / mL) and 1ml of CCTCC M 2017330 bacterial fluid (1×107 CFU / mL), with Sabouraud medium instead of CCTCC M 2017330 bacterial fluid as the control;

[0026] 2) Incubate the inoculated peanut powder in an incubator at 28° C. for 9 days, add 15 mL of 70% methanol water, vortex and put it on a shaker for 30 minutes. Take 3mL supernatant, add 8mL ultrapure water and vortex centrifuge;

[0027] 3) Take 8 mL of the supernatant and use the immunoaffinity column-HPLC method to determine the content of aflatoxin B1 (Table 3), and there are 3 repetitions in the experiment.

[0028] Table 3 Control effect of biocontrol bacteria on peanut aflatoxin

[0029]

[0030] It can be seen from the experimental results that the inhibition rate of CCTCC M 2017330 strain on the toxin production ...

Embodiment 3

[0032] 1) Take 10 grains of Luhua No. 8 peanuts from Anhui peanut field, coat the surface of the peanuts with the fermentation liquid of Enterobacter cloacae 3J1EC, and add 1ml of Aspergillus flavus spore liquid (5×10 5 Individual / mL), with Sabouraud medium instead of CCTCC M 2017330 fermentation broth as a control;

[0033] 2) Cultivate the inoculated peanut grains in an incubator at 28°C for 9 days, then grind the peanut grains into peanut powder, add 15mL of 70% methanol water, vortex and place on a shaker for 30min. Take 3mL supernatant, add 8mL ultrapure water and vortex centrifuge;

[0034] 3) Take 8 mL of the supernatant and use the immunoaffinity column-HPLC method to determine the content of aflatoxin B1 (Table 4), and there are 3 repetitions in the experiment.

[0035] Table 4 Control effect of biocontrol bacteria on peanut Aspergillus flavus

[0036]

[0037] From the experimental results, it can be seen that the inhibition rate of CCTCC M 2017330 strain on the...

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Abstract

The invention belongs to the field of a microorganism, and particularly relates to an enterobacter cloacae biocontrol strain for efficiently inhibiting aspergillus flavus compounded aflatoxin and itsapplication. The enterobacter cloacae biocontrol strain 3J1EC has been preserved in China typical culture preservation center (CCTCC for short) on June 13, 2017; the preservation address is Wuhan University of Wuhan of China; the preservation number is CCTCC No. M2017330. The enterobacter cloacae biocontrol strain can be used for inhibiting aspergillus flavus compounded aflatoxin and preventing the aflatoxin pollution of grain crops.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a biocontrol strain of Enterobacter cloacae capable of efficiently inhibiting the synthesis of aflatoxin by Aspergillus flavus and its application. Background technique [0002] Aspergillus flavus is a pathogenic fungus that can produce a class of strong carcinogenic and highly toxic mycotoxins - aflatoxins, including B, G and M groups, of which B1 is the most common and the most toxic. It can widely pollute peanuts, corn and other food crops, seriously threaten the health of people and livestock, and cause great economic losses. Therefore, it is urgent to strengthen the prevention and control of Aspergillus flavus and toxin pollution. [0003] At present, in the control of Aspergillus flavus, there are three control methods: physical, chemical and biological. However, chemical control is not only costly, but also easy to pollute the environment. While preventing and c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23B9/28C12R1/01A01N63/20
CPCA23B9/28C12N1/20A23V2002/00C12R2001/01C12N1/205A23V2200/10A01N63/20A01N25/14
Inventor 李培武丁小霞王同张奇
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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