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A kind of n-acyl homoserine lactone quenching bacteria and its application in disease prevention and control

An acyl homoserine quenching technology, applied in the field of biological control, can solve the problems of acyl side chain length, saturation difference, etc., and achieve the effects of reducing pesticide abuse, significant biodegradation effect, and significant biocontrol effect

Active Publication Date: 2021-04-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

N-acyl homoserine lactones (N-Acylhomoserine lactones, AHLs) are the unique quorum sensing signals of Gram-negative bacteria. Most of them have the same homoserine lactone ring structure and have acyl side chains, but There are differences in the length and degree of saturation of the acyl side chain

Method used

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  • A kind of n-acyl homoserine lactone quenching bacteria and its application in disease prevention and control
  • A kind of n-acyl homoserine lactone quenching bacteria and its application in disease prevention and control
  • A kind of n-acyl homoserine lactone quenching bacteria and its application in disease prevention and control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The acquisition and identification of embodiment 1 Paleobacterium intermedium bacterial strain D-2

[0048] 1. Isolation and screening of strain D-2

[0049] (1) Soil sample collection: soil samples collected from power plants in Hebei Province as microbial sources

[0050] Soil samples were collected from the power plant in Hebei Province on August 9, 2017. Sampling, bagging, and preservation were carried out as microbial sources and brought back to the school for strain isolation.

[0051] (2) Enrichment culture of bacterial strains: prepare basal salt (MSM) medium, put 20mL MSM medium into a 250mL Erlenmeyer flask for sterilization, add OHHL mother liquor (acetonitrile as solvent) under aseptic conditions after cooling, and make The final concentration of OHHL was 5μM, and 5g soil samples were added at the same time. After culturing for 7 days at 30°C and 200rpm, the inoculum was transferred to the second batch of MSM medium containing 10μM OHHL. After culturing un...

Embodiment 2

[0075] Antibiotic susceptibility analysis of embodiment 2 bacterial strain D-2

[0076] In order to better study the biocontrol potential of the bacterial strain D-2 obtained in Example 1, we conducted in-depth research on the biological characteristics of the bacterial strain. The sensitivity of strain D-2 to different antibiotics was experimentally studied, as Figure 4 shown.

[0077] The experimental results showed that the resistance of the strain D-2 to ampicillin reached 400 μg·mL -1 Above, the resistance to kanamycin reaches 350 μg·mL -1 , the resistance to gentamicin and streptomycin reaches 40 μg·mL -1 , the resistance to chloramphenicol reached 10 μg·mL -1 , no resistance to tetracycline. This result is helpful for selecting appropriate antibiotics as a reference in follow-up studies.

Embodiment 3

[0078] Example 3 Strain D-2 Substrate Spectrum Analysis

[0079] 1. Strain culture and sample collection:

[0080] Under the condition of 30°C, the strain D-2 was activated on the LB solid plate. Pick a single colony and inoculate it into LB liquid medium, and culture overnight at 30°C and 200rpm to obtain a bacterial solution. Take a certain volume (V=1 / OD 600 ) was centrifuged at 4000rpm for 10min, the supernatant was discarded to obtain an OD 600 value bacteria. Will 1 OD 600 Value of the bacteria, inoculated into 1mL MSM inorganic salt medium with different AHLs as the sole carbon source. The reaction mixtures were placed in 2 mL centrifuge tubes, and the centrifuge tubes were incubated at 30° C. and 200 rpm for 24 h. After 24 hours, take 5 μL of the reaction mixture and apply it to the top of a MM agar strip with a width of 1 cm, and then spot the reporter strain CF11 (Agrobacterium tumefaciens NT1) bacteria solution below. Then place the agar strips with the react...

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Abstract

The invention discloses a N ‑Acyl homoserine lactone quencher bacteria and its application in disease control. The present invention found that P. intermedia has quenching activity on various AHLs quorum sensing signal molecules, and isolated a high-efficiency quenching bacterial strain D-2, which can rapidly and significantly degrade various AHLs quorum sensing signal molecules, Including OHHL, OOHL and OdDHL, etc., and can significantly reduce the symptoms of pathogenic bacteria that rely on AHLs to cause disease, and have great application potential in the prevention and treatment of pathogenic bacteria that are mediated by AHLs. Moreover, the activity of the strain is stable, which provides a new development path for a therapeutic strategy that replaces chemical control with biological control and targets blocking quorum sensing without causing selection pressure. It has great potential for popularization and application, and this kind of anti-pathogenic biocontrol bacteria with group quenching as the mode of action has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of biological control. More specifically, it relates to an N-acyl homoserine lactone quenching bacterium and its application in disease prevention and control. Background technique [0002] Pectobacterium carotovorum subsp. carotovora (Pcc) is the main member of the pathogenic bacteria of plant soft rot (Lim J, Jee S, Lee D H, et al..Biocontrol of Pectobacterium carotovorum subsp. carotovorum using bacteria PP1[J ]. Journal of Microbiology and Biotechnology, 2013, 23(8): 1147-1153.). Pcc can not only infect a variety of temperate and subtropical crops and ornamental plants, such as: crops cabbage, radish, tomato, potato, carrot, pepper, celery, cauliflower, etc.; ornamental plants Clivia, lily, violet, narcissus, cactus, etc. Moreover, it can also infect medicinal plants such as unicorn and milk thistle. Bacterial soft rot caused by Pcc has a wide range of occurrences, and has occurred in many countries s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P1/00C12R1/01
CPCA01N63/00C12N1/205C12R2001/01
Inventor 陈少华范兴辉叶田梁梓侨李欣李昊江嘉敏
Owner SOUTH CHINA AGRI UNIV