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Application of bacillus proteolyticus to prevention and treatment of pathogenic bacteria and diseases depending on quorum sensing signal molecules

A quorum sensing signal and Bacillus technology, applied in the field of biological control, can solve the problem that chemical control is not targeted, and achieve the effect of reducing the problem of pesticide abuse

Active Publication Date: 2020-01-10
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The difference between biological control and chemical control such as antibiotics is that chemical control is not targeted and may kill the population itself, while biological control can avoid this

Method used

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  • Application of bacillus proteolyticus to prevention and treatment of pathogenic bacteria and diseases depending on quorum sensing signal molecules
  • Application of bacillus proteolyticus to prevention and treatment of pathogenic bacteria and diseases depending on quorum sensing signal molecules
  • Application of bacillus proteolyticus to prevention and treatment of pathogenic bacteria and diseases depending on quorum sensing signal molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The acquisition and identification of embodiment 1 bacillus strain S-5

[0039] 1. Isolation and screening of strain S-5

[0040] The collected soil samples were screened by enrichment culture with AHLs as the only carbon source. Prepare several bottles of 50mL basic salt MSM medium and put them into a 250mL Erlenmeyer flask, sterilize at 121°C for 20min, add AHLs mother solution under aseptic conditions after cooling, so that the final concentration of AHLs is 2.5μM, weigh 5g soil sample and add MSM In the culture medium, after culturing on a shaker at 30° C. and 200 rpm for 7 days, the inoculation amount (1 mL) of 2% was transferred to the second batch of 50 ml MSM medium with a concentration of AHLs of 5 μM. After culturing for 7 days under the same conditions, they were transferred to 50 mL MSM medium with AHLs concentration of 10 μM according to the inoculum size of 2%, and continued culturing for 7 days. Inoculate by analogy until the concentration of AHLs incre...

Embodiment 2

[0051] Antibiotic susceptibility analysis of embodiment 2 quenching bacterial strain S-5

[0052] In order to better study the biocontrol potential of the bacterial strain S-5 obtained in Example 1, we conducted in-depth research on the biological characteristics of the bacterial strain. The sensitivity of strain S-5 to different antibiotics was experimentally studied as follows:

[0053] Take the strain out of the -80°C refrigerator, activate the quenched bacteria with a solid LB plate, pick a single colony and inoculate it into 1mL liquid LB, and cultivate to OD 600 The value is between 1.0-1.5, and the bacterial liquid is used as the inoculum. With 5 mL liquid LB medium as the system, 6 kinds of antibiotics including kanamycin, ampicillin, gentamycin, streptomycin, carbenicillin and tetracycline were added to test the antibiotic susceptibility of quenched bacteria. In addition to the control group, 13 concentrations of 5 μg mL were also set for each antibiotic. -1 , 10μg...

Embodiment 3

[0055] Example 3 Determination of Quenching Bacterial Strain S-5 Growth and Degradation AHLs Relationship Curve

[0056] Take the strain out of the -80°C freezer, activate the strain with a solid LB plate, pick a single colony and add it to five 50mL centrifuge tubes containing 5mL of liquid LB, and after culturing overnight, take the bacterial solution (V=1 / OD 600 ) into a sterilized 2mL centrifuge tube, centrifuge at 10000rpm for 1min, and discard the supernatant. After the precipitated bacteria were resuspended with the solution in the sterilized MSM medium, they were inoculated into 20 mL of MSM medium, and AHLs mother solution (concentration: 100 mmol / L) was added to make the final concentration 0.4 mM. Cultivate at 30°C and 200rpm, collect samples at 24h and 48h time points and measure OD 600 The value indicates the growth of the quenched bacteria, and the residual amount of AHLs determined by HPLC indicates the degradation of AHL by the quenched bacteria.

[0057] T...

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Abstract

The invention discloses an application of bacillus proteolyticus in prevention and treatment of pathogenic bacteria and diseases depending on quorum sensing signal molecules. The research finds that the bacillus proteolyticus has a very good effect of degrading dependent quorum sensing signal molecules, an efficient quenching strain of the microorganism quorum sensing signal molecules is screened,which is the bacillus proteolyticus strain S-5, and is preserved in the Guangdong Microbial Culture Collection Center on September 4, 2019, and a preservation number of the bacillus proteolyticus strain S-5 is GDMCC No. 60764. The strain has the activity of quenching AHLs and DSF quorum sensing signal molecules, the quenching effect is stable and remarkable, so that the anti-pathogenicity drug has huge application potential in the aspect of preventing and treating pathogenic bacteria harm caused by AHLs and DSF quorum sensing signals, and the anti-pathogenicity drug taking quorum quenching asan action mode has wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of biological control. More specifically, it relates to the application of Bacillus proteolyticus in the prevention and treatment of pathogenic bacteria and diseases that depend on quorum sensing signal molecules. Background technique [0002] Homeostasis is the state of a stable ecosystem, which is changing, not static. When the external environmental conditions do not change drastically, the number of each biological population will fluctuate at a certain level, forming a relatively balanced state (Peng Haowen, Li Qiqin, Lin Wei. Overview of biological control research and its application[J] . Guangxi Agricultural Biosciences, 2004, 2:170-174.). The outbreak of pests and diseases is the performance when the relative balance of the ecosystem is broken, and this phenomenon is most obvious in the agricultural ecosystem. [0003] Every year, the occurrence of diseases and insect pests brings huge economic l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 李绮婷陈少华范兴辉叶田李欣梁梓侨
Owner SOUTH CHINA AGRI UNIV
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