Application of Pseudomonas nitroreductors hs-18 in the prevention and treatment of pathogenic bacteria mediated by ahls

A technology of Pseudomonas and C6-HSL, which is applied in the fields of application, chemicals for biological control, botany equipment and methods, etc., can solve the problems that are rarely studied, achieve good degradation activity, and the degradation effect is efficient and significant Effect

Active Publication Date: 2021-10-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on quorum quenching mainly focuses on the quenching of specific types of quorum sensing signals, and there is little research on the quenching ability of quenching bacteria.

Method used

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  • Application of Pseudomonas nitroreductors hs-18 in the prevention and treatment of pathogenic bacteria mediated by ahls
  • Application of Pseudomonas nitroreductors hs-18 in the prevention and treatment of pathogenic bacteria mediated by ahls
  • Application of Pseudomonas nitroreductors hs-18 in the prevention and treatment of pathogenic bacteria mediated by ahls

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Degradation system setting of Pseudomonas nitroreducens HS-18 to AHLs

[0033] First, use LB liquid culture based on 30 ℃, 200rpm constant temperature shaker overnight culture strain HS-18 to obtain OD 600 =0.5 seed solution, take 300 μl seed solution and add the same volume of fresh LB liquid medium, and add MOPS with a final concentration of 50 mM and AHLs with different carbon chain lengths and different substituents at a final concentration of 10-50 μM to prepare a degradation system (The appropriate concentration of different AHLs signals is selected according to the different intensities of color development of the reporter strain), and cultured at 30°C, 200rpm in a constant temperature shaker for 12h, 24h, and 36h, and the group without adding bacterial suspension was used as a control. The signals of exogenously added AHLs were extracted with the same volume of ethyl acetate as that of the degradation system.

[0034] Table 1 lists the AHLs used to ex...

Embodiment 2

[0038] Example 2 Quantitative detection of short-chain AHLs using reporter strain Violet bacteria (Chromobacterium violaceum) CV026

[0039] Violet bacteria CV026 was activated on LB solid plates, and CV026 was cultured overnight in LB liquid medium. Prepare LB solid plates and cut into 0.8 cm wide strips of interphase agar. Take 10 μl of the ethyl acetate extract from the degradation system described in Example 1 and spot it on the top of the LB agar strip, and spot a row of reporter strain droplets of similar size below the sample loading place. After the samples and the reporter strains were dripped dry, the plates were incubated in a constant temperature incubator at 28°C for 16 hours. The area where AHLs diffused could induce violaceum CV026 to produce violacein and make the cells appear purple. The distance of the purple reporter strain CV026 is proportional to the content of AHLs to be detected. The result is as figure 1 It shows that the bacterial strain HS-18 in t...

Embodiment 3

[0040] Example 3 Quantitative detection of medium and long chain AHLs using reporter strain Agrobacterium tumefaciens NT1 (traR, tra::lacZ749)

[0041] Agrobacterium tumefaciens NT1 was activated on LB solid plates, and NT1 was cultured overnight in LB liquid medium containing kanamycin at a final concentration of 50 μg / ml. Prepare MM solid plates with X-gal (5-bromo-4-chloro-3-indole-β-D-galactopyranoside) added at a final concentration of 40 μg / ml and cut into 0.8 cm wide interphase agar strips . Take 10 μl of the ethyl acetate extract from the degradation system described in Example 1 and spot it on the top of the MM agar strip, and spot a row of reporter strain droplets of similar size below the sample loading place. After the samples and the reporter strains were dripped dry, the plates were wrapped in tin foil and placed in a constant temperature incubator at 28°C in the dark for 16 hours. The area where AHLs spread can induce Agrobacterium tumefaciens NT1 to produce β...

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Abstract

The present invention belongs to the technical field of molecular biological control. The present invention finds that the nitroreductive Pseudomonas strain HS-18 has good degradation activity to the AHLs with C4-C14 chain length used for detection, and the degradation effect is highly efficient and significant. A new biocontrol agent is provided for the biological control of pathogenic bacteria mediated by AHLs, thereby broadening the types of group quenching bacteria of AHLs signaling molecules.

Description

technical field [0001] The invention relates to the technical field of molecular biological control, and more specifically relates to the application of nitroreductor pseudomonas HS-18 in the prevention and treatment of AHLs-mediated disease-causing pathogens. Background technique [0002] Quorum sensing (quorum sensing, QS) is a communication mechanism between cells, and quorum sensing signal is the "language" to complete this communication mechanism. Quorum sensing signal is a small molecular compound synthesized and secreted by bacteria. The production of this small molecular compound runs through the entire cell growth cycle and can freely diffuse or actively transport to the extracellular environment. When the number of cells increases, the concentration of extracellular quorum sensing signals also increases, and cells sense the number of cells in the environment by sensing the concentration of quorum sensing signals. When the concentration of the quorum sensing signal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N63/27A01P1/00
CPCA01N63/27
Inventor 张炼辉王惠杉廖立胜范兴辉
Owner SOUTH CHINA AGRI UNIV
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