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A strain of Bacillus amyloliquefaciens, fermentation product, preparation method and application thereof

A technology of amylolytic spores and fermentation products, which is applied in the field of biotechnology, can solve problems such as the lack of mechanism of occurrence, and achieve the effect of simple preparation method, easy operation, and strong antibacterial potential

Active Publication Date: 2020-12-11
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In actual production, in addition to disease-resistant varieties, the prevention and control of rice base rot should still be assisted by control chemicals. Since this disease was a new disease first reported in the late 1970s, the disease is currently increasing and spreading year by year. However, there is still a lack of more in-depth and systematic research on the mechanism of the disease at home and abroad.

Method used

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  • A strain of Bacillus amyloliquefaciens, fermentation product, preparation method and application thereof
  • A strain of Bacillus amyloliquefaciens, fermentation product, preparation method and application thereof
  • A strain of Bacillus amyloliquefaciens, fermentation product, preparation method and application thereof

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Effect test

Embodiment 1

[0051] Example 1 Screening and identification of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) E3 strain

[0052] 1. Screening of strains

[0053] (1) Preliminary screening of Bacillus amyloliquefaciens: Take 1 g of a citrus orchard soil in Hunan Province and add it to 10 mL of sterile water, incubate at 200 rpm for 30 min, then dilute with water, the dilution gradient is 10×, 100× and 1000×, take 100 μL of each Dilutions of rice root rot bacteria (hereinafter referred to as EC1, the full-length genome of this strain has been published on NCBI GenBank, the sequence number is NCBI Reference Sequence: NZ_CP006929, and the website is https: / / www.ncbi.nlm.nih .gov / nuccore / NZ_CP006929.1) on LB solid medium. Preparation of LB solid medium containing EC1: use LB liquid medium to cultivate EC1 bacteria, cultivate to OD at 28°C, 200rpm 600 It is about 1, then mixed at 40-50°C at a ratio of 10 μL of bacterial suspension per ml of LB medium, poured about 10mL of medium into e...

Embodiment 2

[0081] (1) Preparation of fermentation stock solution

[0082] Prepare 100mL LB liquid culture medium, inoculate the biocontrol bacterium (ie E3 bacterium) that embodiment 1 obtains according to the inoculum amount of 1% (volume ratio), 28 ℃, 200rpm, cultivate for three days and obtain fermentation stock solution.

[0083] (2) Preparation of EC1 bacterial solution:

[0084] Inoculate EC1 single colony LB liquid medium, culture at 28°C, 200rpm, grow to OD 600 = about 0.6, centrifuge at 5000rpm for 10 minutes to collect the bacterial cells, suspend them with sterile water, and use the colony plate counting method to dilute to obtain different concentrations of EC1 bacterial solutions, which are 1.4×10 7 cfu / mL, 1.4×10 6 cfu / mL, 1.4×10 5 cfu / mL, 1.4×10 4 cfu / mL, 1.4×10 3 cfu / mL;

[0085] (3) Disease prevention and growth promotion characteristics of Bacillus amyloliquefaciens E3 on rice seeds

[0086] In order to test the antibacterial effect of biocontrol bacteria E3 on r...

Embodiment 3

[0089] The antibacterial spectrum test of embodiment 3 bacillus amyloliquefaciens E3 crude lipopeptide

[0090] 1. Antibacterial lipopeptide extraction

[0091] The lipopeptide was extracted by HCl precipitation combined with organic solvent extraction. Take 100 mL of E3 fermentation stock solution, centrifuge at 8000 rpm for 15 minutes, discard the precipitate, slowly add HCl to the fermentation supernatant while stirring, adjust the pH value to 2.0, and let it stand overnight at 4°C. The precipitated protein was centrifuged at 8000r / min for 15min, the supernatant and precipitate after salting out the protein were collected, and the precipitate was extracted with 5 times the volume of acetone. After the extraction, centrifuge at 8000r / min for 15min, collect the supernatant, extract the precipitate again with the same organic solvent, centrifuge, and collect the supernatant extract. Air-dry the two extracts in a fume hood to obtain a crude extract, and collect and store the ...

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Abstract

The invention discloses a strain of bacillus amyloliquefaciens, a fermentation product, a preparation method and application thereof. The strain is named Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) E3, which was deposited in the China Center for Type Culture Collection (CCTCC) located in Wuhan University, Wuhan, China on December 7, 2017, and the preservation number is CCTCC NO: M 2017768; the strain It can effectively inhibit the infection of rice seeds by rice base rot pathogens, increase the germination rate, and can be used as a biocontrol bacteria for the prevention and treatment of rice base rot. The invention also provides a preparation method of the fermentation stock solution of the bacterial strain, which is simple and easy to operate. The present invention also provides the antibacterial lipopeptide of the bacterial strain and its extraction method. The antibacterial lipopeptide exhibits inhibitory effect on various plant pathogenic bacteria, has a wide antibacterial spectrum, and can be used to develop high-efficiency, broad-spectrum, low-toxicity, Environmentally friendly microbial agents.

Description

technical field [0001] The invention belongs to the field of biological high technology, and in particular relates to a strain of bacillus amyloliquefaciens, a fermentation product and a preparation method and application thereof. Background technique [0002] In the process of agricultural production, plant pathogenic bacteria cause plant diseases that seriously affect the growth and yield of plants, and even cause plant death. At present, the control of plant diseases mainly relies on chemical pesticides, which have made great contributions to the safe production of agricultural products, but the widespread use of chemical pesticides and the increase in the amount of pesticides have caused a series of problems, which have increased the resistance of certain pathogenic bacteria, and at the same time It will cause environmental pollution, so the development of biological pesticides is imminent, and the use of microorganisms to inhibit plant pathogenic bacteria provides the p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/00C07K1/14A01N63/22A01N63/50A01P1/00C12R1/07
CPCA01N63/00A01N63/10C07K1/145C12N1/20C12P21/00C12N1/205C12R2001/07
Inventor 徐领会梁丽琼王鑫王俊霞王庆伟杨小贝陈小云
Owner SOUTH CHINA AGRI UNIV
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