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N-acyl high serine lactone quenching bacterium and application thereof in disease control

A quenching and bacterial strain technology, which is applied in N-acyl homoserine lactone quenching bacteria and its application in disease prevention and control, can solve the problems of acyl side chain length and saturation difference, and reduce the problem of pesticide abuse Effect

Active Publication Date: 2019-01-11
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 -Acyl homoserine lactones, AHLs) are quorum sensing signals unique to Gram-negative bacteria. Most of them have the same homoserine lactone ring structure and acyl side chains, but the length and saturation of the acyl side chains are different.

Method used

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  • N-acyl high serine lactone quenching bacterium and application thereof in disease control
  • N-acyl high serine lactone quenching bacterium and application thereof in disease control
  • N-acyl high serine lactone quenching bacterium and application thereof in disease control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Acquisition and Identification of Paleobacterium intermedium 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 strains: prepare basic salt (MSM) medium, put 20 mL of MSM medium into a 250 mL Erlenmeyer flask for sterilization, add OHHL mother solution under sterile conditions after cooling (acetonitrile as solvent) , so that the final concentration of OHHL was 5 μM, and 5 g of soil samples were added at the same time. After culturing for 7 days at 30 °C and 200 rpm, the inoculum was transferred to the second batch of MSM culture containing 10 μM OHHL Base. After culturing under t...

Embodiment 2

[0072] Antibiotic susceptibility analysis of embodiment 2 bacterial strain D-2

[0073] 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.

[0074] 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 reached 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

[0075] Example 3 Analysis of strain D-2 substrate spectrum

[0076] 1. Strain culture and sample collection:

[0077] Under the condition of 30 ℃, 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 200 rpm to obtain a bacterial liquid. Take a certain volume (V=1 / OD 600 ) was centrifuged at 4000 rpm for 10 min, and 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 h, take 5 μL of the reaction mixture and spot it on the top of a 1 cm wide MM agar strip, and then spot the reporter strain CF11 ( Agrobacterium tumefaciens NT1) bacteria solution. Then place the agar strips with the reaction mixture...

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Abstract

The invention discloses an N-acyl high serine lactone quenching bacterium and an application thereof in disease control. Researches show that an ochrobactrum intermedium has quenching activity to various AHLs quorum sensing signal molecules, is separated to obtain an efficient quenching bacterium strain D-2, can degrade the various AHLs quorum sensing signal molecules quickly and remarkably, including OHHL, OOHL and OdDHL and the like, can alleviate AHLs dependent pathogenic bacteria disease symptoms remarkably and has a huge application potential in preventing and treating AHLs-mediated pathogenic bacterium harm. The strain is stable in activity, provides a novel development path of a treatment strategy for replacing chemical prevention with bioprevention and blocking quorum sensing as atarget without inducing selective pressure and has a huge popularizing and applying potential in preventing and treating quorum sensing mediated pathogenic bacteria harm. The anti-pathogenic biocontrol bacterium acting as quorum sensing has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of biological control. More specifically, a N -Acyl homoserine lactone quencher bacteria and its application in disease control. Background technique [0002] Pectinobacillus carotovora subsp. Pectobacterium carotovorum subsp . carotovora , Pcc) are the main members of plant soft rot pathogens (Lim J, Jee S, Lee D H, et al..Biocontrol of Pectobacterium carotovorum subsp. carotovorum usingbacteriophage 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 countrie...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/01
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
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