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A non-toxic Aspergillus flavus hubxy33 and its application

A technology of Aspergillus flavus and aflatoxin, which is applied in the field of microorganisms, can solve the problems of mycotoxin pollution, etc., and achieve the effects of high safety, good inhibition effect and strong competitive advantage

Active Publication Date: 2022-03-11
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

Existing reports are used as non-toxin-producing Aspergillus flavus and uses thereof (such as CN 110157626 A, CN 110305796 A, CN 110129212 A, CN 107177516, CN104789480 A) for biological control, only for not producing aflatoxin, it may be possible after field application Risk of contamination with mycotoxins such as aflatoxin, versicolor and other mycotoxins

Method used

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  • A non-toxic Aspergillus flavus hubxy33 and its application
  • A non-toxic Aspergillus flavus hubxy33 and its application
  • A non-toxic Aspergillus flavus hubxy33 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Separation and purification

[0045] DG-18 medium (article number: HB0296, Qingdao Hi-Tech Industrial Park Haibo Biotechnology Co., Ltd.) was selected as the medium for the isolation of Aspergillus flavus. The soil samples collected from the peanut planting field in Xiangyang, Hubei were treated by removing plant residues and large-grained stones, soil grinding, and mixing.

[0046] Weigh 10.0g of the ground soil sample and add it into 90mL of sterilized water, put it into a shaker and shake for 2h to make a 100ml sample suspension. Take 50 μl of the suspension, add it to the DG-18 medium plate, and spread it evenly with a coating rod. Then put the coated plate into a constant temperature incubator and culture it at 28±1° C., 90% relative humidity and dark conditions.

[0047] After 5 days, the bacterium colonies with yellow-green spores in the appearance were picked and transferred to a new DG-18 medium plate for secondary streaking and purification until a single...

Embodiment 2

[0070] 1. Strain toxin-producing culture

[0071] The non-toxigenic Aspergillus flavus strain HuBXY33 to be tested was inoculated on DG-18 solid medium for activation. After culturing at 28±1° C. for 5 days in the dark, the aspergillus flavus conidia on the plate were washed with sterilized 0.1% Tween 80 to obtain a conidia suspension.

[0072] Determine the conidia suspension concentration using a hemocytometer.

[0073] Take a certain amount of conidia suspension and inject it into the Erlenmeyer flask filled with sterilized 30ml liquid Sabouraud medium, so that the final concentration is 4×10 5 pieces / ml. Place the Erlenmeyer flask on a shaker at 28±1°C, 200rpm in the dark.

[0074] 2. Determination of strain toxin content

[0075] After 5 days, use sterilized gauze to filter the mycelial culture solution, discard the mycelium ball, collect the toxin-producing culture solution and store it in a centrifuge tube, let it stand for 1 hour, pipette 500μl of the toxin-produci...

Embodiment 3

[0083] 1. Preparation of conidia suspension

[0084] The non-toxigenic Aspergillus flavus strain HuBXY33 to be tested and the toxigenic Aspergillus flavus standard strain 3.4408 and the toxin-producing Aspergillus flavus (Aspergillus flavus) isolated from different regions are numbered (SCPA-32-12 (Peng'an, Sichuan) , XZCY-24-6 (Zayu, Tibet), LNFX-25-1 (Fuxin, Liaoning), HBHA-129-1 (Hong’an, Hubei), HBYL-12-7 (Yangluo, Hubei), HBTS-94-2 ( Hebei Tangshan), JXZS-118-9 (Jiangxi camphor tree) and JXZS-69-3 (Jiangxi camphor tree) were respectively inoculated in DG-18 solid medium for activation. After culturing for 5 days at 28±1°C in the dark, use sterilized The conidia of Aspergillus flavus on the plate were washed with 0.1% Tween 80 and placed in a 10 ml centrifuge tube to obtain conidia suspension.

[0085] Determine the concentration of the conidia suspension with a hemocytometer under a microscope, and dilute the concentration of conidia to 1×10 with sterile water. 6 pcs / ml...

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Abstract

The invention provides a non-toxin-producing Aspergillus flavus HuBXY33 and its application, and particularly relates to the technical field of microorganisms. The invention provides a non-toxin-producing Aspergillus flavus HuBXY33, and the preservation number of the Aspergillus flavus HuBXY33 is CCTCC NO: M 2020519. The Aspergillus flavus HuBXY33 of the present invention does not produce aflatoxin, aflatoxin and aspergillus versicolor, is a biocontrol strain with high safety and strong reproductive ability, and has efficient competitive inhibition of the growth of toxin-producing Aspergillus flavus, Reproductive and toxin-producing ability, can significantly reduce the content of aflatoxin, aflatoxin and versicolor in agricultural products such as peanuts.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a non-toxin-producing Aspergillus flavus HuBXY33 and its application. Background technique [0002] After Aspergillus flavus infects crops, it can produce toxic secondary metabolites such as Aflatoxins (AFT), Aflatrem, and Sterigmatocystin (ST), which seriously threaten consumer safety and People's lives and health. Aflatoxins have acute and chronic toxicity, can cause cancer, teratogenicity, and mutagenesis. They are the most toxic and most carcinogenic mycotoxins found in contaminated agricultural products so far. Aflatoxin is an indole diterpene mycotoxin with tumorigenic properties and acute neurotoxic effects, causing tremors in animals. Aspergillus versicolor is an intermediate product in the late stage of the synthesis of aflatoxin by Aspergillus flavus. Because of its similar structure to aflatoxin, its toxicity and carcinogenicity have been highly valued by countries all ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01G7/06A01G22/20A01G22/40C12R1/67
CPCA01G7/06A01G22/20A01G22/40
Inventor 李培武张奇岳晓凤白艺珍马飞荆丹
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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