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AHLs quenching bacterium and application thereof in control of pathogenic bacteria depending on AHLs

A technology of pathogenic bacteria and strains, applied in the application field of pathogenic bacteria, can solve problems such as microbial degradation, achieve good degradation activity, stable degradation effect, and solve the effects of pesticide or antibiotic abuse and drug resistance

Active Publication Date: 2021-11-26
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the risk of degradation of microbial bacteria, it is of great significance to continuously enrich the bio-control bacteria library

Method used

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  • AHLs quenching bacterium and application thereof in control of pathogenic bacteria depending on AHLs
  • AHLs quenching bacterium and application thereof in control of pathogenic bacteria depending on AHLs
  • AHLs quenching bacterium and application thereof in control of pathogenic bacteria depending on AHLs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Isolation and screening of embodiment 1 Rhodococcus bacterial strain SXT-7

[0049] 1. Soil sample collection:

[0050] Several soil samples were collected from the offshore shoals of mangrove forests in Shenzhen. After the samples were retrieved, they were stored in a 4°C freezer for later use.

[0051] 2. Enrichment culture of bacterial strains:

[0052] Soil samples collected from mangrove parks were used as microbial sources, and AHLs were used as the only carbon source to enrich and screen the microorganisms. Put the prepared 50mL basal salt medium (MSM) into a 250mL Erlenmeyer flask and sterilize it at 121°C for 20min. After cooling, add N-(3-oxohexanoyl)-L - Homoserine lactone (3OC6HSL) mother solution (concentration is 100mmol / L, solvent is methanol), so that the final concentration of bacterial quorum sensing signal molecule 3OC6HSL is 5 μmol / L. At the same time, 5 g of microbial source soil samples were added, placed on a shaker at 30°C and 200 r / min for 7 ...

Embodiment 2

[0059] The identification of embodiment 2 Rhodococcus strain SXT-7

[0060] In this example, morphological identification, 16S rDNA phylogenetic analysis, and physiological and biochemical identification were carried out on the quenched bacterium SXT-7, and the strain was identified as Rhodococcus sp. details as follows:

[0061] 1. Morphological characteristics of the colony: The colony of the quenched strain SXT-7 is pale yellow and round in the LB solid plate, the colony is smooth and shiny, with neat edges, and the surface is viscous and translucent. like figure 1 As shown; when growing in LB liquid medium, it was diffuse and turbid, and grew well at 28°C.

[0062] 2. Morphological characteristics of bacteria: such as figure 2 As shown, the bacterial cells are rod-shaped, without flagella, and have mucus on the surface.

[0063] 3. 16S rDNA identification and phylogenetic analysis: Using the genome of strain SXT-7 as a template, PCR amplification was performed using b...

Embodiment 3

[0066] Example 3 Detection of degradation activity of Rhodococcus strain SXT-7 on AHLs signal

[0067] In this example, the reporter strain Agrobacterium tumefaciens CF11 was used to detect the degradation effect of strain SXT-7 on AHLs. The strain SXT-7 was activated with LB solid medium plate, and the plate was placed in an incubator at 28°C for 48 hours. Pick a single colony and inoculate it into liquid LB medium, and culture overnight at 28°C and 200rpm to obtain a bacterial solution. Obtain SXT-7 cells with an OD6oo value of 1.0 and inoculate them into 1 mL of MSM inorganic salt medium. MSM only contains AHLs as the sole carbon source. The concentration of AHLs in the reaction system was 40μmol / L. The reaction mixture is incubated under suitable conditions. Then take the reaction mixture and the bacterial liquid of the reporter strain Agrobacterium tumefaciens CF11 for spotting. At the same time, Bacillus thuringiensis subsp. israelensis B23 known to be capable of que...

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Abstract

The invention belongs to the technical field of agricultural disease control, and particularly relates to an AHLs quenching bacterium and application thereof in control of pathogenic bacteria depending on AHLs. The invention discloses Rhodococcus sp. SXT-7 capable of degrading quorum sensing signal molecules AHLs. The strain was preserved in Guangdong Microbial Culture Collection Center on July 21, 2021, and the preservation number is GDMCC No.61818. The research indicates that the Rhodococcus sp. SXT-7 has favorable degradation activity for quorum sensing signal molecules AHLs, and has stable and obvious degradation effect; the degradation rate of the strain SXT-7 for bacterial quorum sensing signal molecules is positively correlated with the growth of the strain SXT-7; and the strain SXT-7 has obvious biological control effect on controlling AHLs-mediated pathogenic bacteria. The Rhodococcus sp. SXT-7 has huge application potential in the aspects of degrading quorum sensing signal molecules AHLs and controlling AHLs-mediated pathogenic bacteria hazards, thereby providing a new way and method for biological control of diseases.

Description

technical field [0001] The invention belongs to the technical field of agricultural disease prevention and control. More specifically, it relates to an AHLs-quenching bacterium and its application in the prevention and treatment of AHLs-dependent pathogenic bacteria. Background technique [0002] Diseases caused by bacteria are a major problem in agriculture. In the context of green agriculture, the use of chemical pesticides is gradually restricted. Therefore, green, efficient and simple new disease control strategies have become research hotspots in recent years. [0003] Quorum sensing (QS) is a phenomenon in which bacterial cells communicate with each other through signal molecules. Bacterial cells secrete and sense the concentration of specific signal molecules. When the concentration of signal molecules reaches a certain threshold, bacterial cells will activate specific genes, especially It is the expression of many disease-causing genes, which provides a new way of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P1/00A01G13/00C12R1/01
CPCA01N63/20A01G13/00
Inventor 周筱帆陈文娟罗青青
Owner SOUTH CHINA AGRI UNIV
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