Method for extracting natural compounds from cell-free supernatant of Burkholderia and application thereof in controlling wheat scab

A natural compound, the technology of Burkholderia, is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., which can solve the problems of slow drug update, increased difficulty in prevention and control, and high cost, and achieve strong in vitro antibacterial activity, Good promotion prospect, strong bactericidal effect

Inactive Publication Date: 2019-09-27
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these agents also have certain problems. Some studies have shown that the target β-tubulin of benzimidazole fungicides will undergo point mutations, and the three-dimensional conformation of the protein will change accordingly, and the affinity between pathogenic bacteria and agents will decrease.
Moreover, due to long-term large-scale unreasonable use, slow update of agents, high cost, and increasing drug resistance of pathogenic bacteria, it is more difficult to prevent and control, causing environmental pollution
[0006] In view of the fact that there are no wheat varieties with high resistance to head blight and the lack of high-efficiency and low-toxicity chemical fungicides, biological control has extensive practical significance

Method used

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  • Method for extracting natural compounds from cell-free supernatant of Burkholderia and application thereof in controlling wheat scab
  • Method for extracting natural compounds from cell-free supernatant of Burkholderia and application thereof in controlling wheat scab
  • Method for extracting natural compounds from cell-free supernatant of Burkholderia and application thereof in controlling wheat scab

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) Bacterial culture: Pick a single colony of Burkholderia on LB agar plate and transfer it to LB liquid medium in a 5ml test tube. The temperature is 30°C, and the shaking rate is 200rpm for overnight culture;

[0045] (2) Collection of supernatant: centrifuge the bacterial suspension obtained in step (1) at 8000 rpm for 10 minutes and collect the supernatant to obtain a cell-free supernatant (CFCS);

[0046] (3) Extraction and evaporation: extract the above cell-free supernatant with an equal volume of ethyl acetate solution, use a separatory funnel to collect the ethyl acetate layer, and centrifuge the obtained ethyl acetate extract at 5000rpm for 5min and filter it through a 0.22 micron organic filter After filtering, use a rotary evaporator to evaporate, vacuum degree 0MPa, temperature 45°C, rotary evaporation to obtain a solid substance;

[0047] (4) Separation and purification: dissolve the above-mentioned solid matter with ethyl acetate solution, and the obtai...

Embodiment 2

[0051] (1) Bacterial culture: Pick a single colony of Burkholderia on LB agar plate and transfer it to LB liquid medium in a 5ml test tube. The temperature is 30°C, and the shaking rate is 200rpm for overnight culture;

[0052] (2) Collection of supernatant: centrifuge the bacterial suspension obtained in step (1) at 9000 rpm for 10 minutes and collect the supernatant to obtain a cell-free supernatant (CFCS);

[0053] (3) Extraction and evaporation: extract the above cell-free supernatant with an equal volume of ethyl acetate solution, collect the ethyl acetate layer with a separatory funnel, and then centrifuge the obtained ethyl acetate extract at 6000rpm for 5min and filter with a 0.22 micron organic filter Finally, use a rotary evaporator with a vacuum of 0 MPa and a temperature of 45°C to obtain a solid substance by rotary evaporation;

[0054] (4) Separation and purification: dissolve the above-mentioned solid matter with ethyl acetate solution, and the solution obtaine...

Embodiment 3

[0056] (1) Bacterial culture: Pick a single colony of Burkholderia on LB agar plate and transfer it to LB liquid medium in a 5ml test tube. The temperature is 30°C, and the shaking rate is 200rpm for overnight culture;

[0057] (2) Collection of supernatant: centrifuge the bacterial suspension obtained in step (1) at 10,000 rpm for 10 minutes and collect the supernatant to obtain a cell-free supernatant (CFCS);

[0058] (3) Extraction and evaporation: extract the above cell-free supernatant with an equal volume of ethyl acetate solution, collect the ethyl acetate layer with a separatory funnel, and then centrifuge the obtained ethyl acetate extract at 7000rpm for 5min and filter with a 0.22 micron organic filter Finally, use a rotary evaporator with a vacuum of 0 MPa and a temperature of 45°C to obtain a solid substance by rotary evaporation;

[0059] (4) Separation and purification: Dissolve the above-mentioned solid matter with ethyl acetate solution, and the obtained solut...

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Abstract

The invention discloses a method for extracting natural compounds from cell-free supernatant of Burkholderia, which comprises the following steps: 1) culturing Burkholderia R456 in LB liquid medium, collecting bacterial suspension, collecting the supernatant by centrifugation and obtaining a cell-free supernatant; 2) extracting the cell-free supernatant with an ethyl acetate solution to obtain an ethyl acetate extract, and then treating the extract to obtain a solid substance; 3) dissolving the solid substance with an ethyl acetate solution, treating the obtained solution and obtaining the active natural compound as shown in the formula below by the preparative liquid chromatography separation. The invention also discloses the application of the above-mentioned Burkholderia R456 and natural compounds in controlling wheat scab. The natural active compound prepared by the invention has stable structure, high purity and good bactericidal activity, can be used as a new control agent for controlling wheat scab, and has a good promotion prospect in the field of agricultural production.

Description

technical field [0001] The invention belongs to the field of extraction and preparation of natural active compounds, and in particular relates to a method for extracting natural active compounds from Burkholderia cell-free supernatant and the application of the Burkholderia and natural active compounds in preventing and treating wheat scab . Background technique [0002] Wheat scab (Fusarium head blight) is a worldwide epidemic disease. my country is one of the countries with the largest affected area in the world. Its causative agent is Fusarium graminearum (Fusarium graminearum). From 2011 to 2015, the average annual occurrence area of ​​wheat scab in my country exceeded 80 million mu, accounting for about 25% of the total planting area. The incidence was particularly severe in 2012, with an infected area of ​​nearly 150 million mu and an economic loss of US$1.5 billion. Wheat scab can not only cause serious yield loss, but also produce a variety of mycotoxins, which seri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D487/04A01P3/00C12P17/18C12R1/38
CPCC07D487/04C12P17/182
Inventor 李斌刘柳张洁净范香虹陈云尹燕妮王晓璇张牧晨
Owner ZHEJIANG UNIV
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