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Quorum sensing signal molecule DSF (Diffusible Signal Factor) quenching strain and application thereof

A quorum sensing signal and signal technology, which is applied in the direction of applications, bacteria, and chemicals for biological control, can solve problems such as health and ecosystem hazards, achieve environmental friendliness, stable degradation performance, and reduce antibiotic abuse.

Active Publication Date: 2018-10-02
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are a large number of microorganisms that can degrade signal molecules in nature. There are many types, large numbers, and easy to cultivate. In addition, the wide and continuous use of antibiotics in the prevention and control of plant diseases has caused more and more pathogenic bacteria to develop drug resistance, which will harm human health. Potentially huge harm to the ecosystem

Method used

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  • Quorum sensing signal molecule DSF (Diffusible Signal Factor) quenching strain and application thereof
  • Quorum sensing signal molecule DSF (Diffusible Signal Factor) quenching strain and application thereof
  • Quorum sensing signal molecule DSF (Diffusible Signal Factor) quenching strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Isolation and screening of Ralstonia skinneri strain F20

[0041] 1. Isolation and purification of bacterial strains

[0042] (1) Soil sample collection: The sweet potato field soil cultivated for many years was collected as the microbial source.

[0043] The soil samples were collected on March 16, 2017 from the sweet potato field soil cultivated all year round in Nanhai District and Shunlu Gangchang, Foshan City, Guangdong Province. The soil from the surface to the depth of 5 cm was sampled, bagged, and preserved as a microbial source for strain isolation.

[0044] (2) Enrichment culture of strains: Prepare MSM medium, put 50 mL of MSM medium into a 250 mL Erlenmeyer flask for sterilization, add DSF mother solution under aseptic conditions after cooling (the concentration of the mother solution is 100 mM, methanol is solvent), so that the final mass concentration of DSF was 0.01 mM, and at the same time, 5 g of soil samples were added, and after 7 days of i...

Embodiment 2

[0055] Example 2 Identification of Ralstonia skinneri strain F20

[0056] 1. Morphological identification of strain F20

[0057] (1) Colony morphological characteristics: Streak the above strain F20 on LB solid medium and culture at 30°C for 48 h. Such as figure 1 As shown, the color of the colony is beige, and the colony is slightly raised, indicating that it is smooth and opaque, with neat edges. Strain F20 was diffusely turbid and aerobic in LB broth.

[0058] (2) Cell morphology characteristics: such as figure 2 As shown, the cells are spherical, or short rod-shaped, with a size of (0.5-1.4)×(0.4-0.7) μm.

[0059] 2. Phylogenetic analysis of strain F20

[0060] 16S rDNA sequence and phylogenetic analysis: The 16S rDNA gene sequence of the strain F20 was obtained, with a length of 1416bp, and compared with the NCBI database (http: / / www.ncbi.nlm.nih.gov / ), it was found that the strain F20 and Ralstonia pickettiiDSM 6297T (LN681565.1) has very good homology, with a...

Embodiment 3

[0062] Example 3 Antibiotic susceptibility analysis of bacterial strain F20

[0063] In order to better study the biocontrol potential of the bacterial strain F20 obtained in Examples 1 and 2, we studied the antibiotic sensitivity of the bacterial strain F20. The results are shown in Fig. 4, the resistance of the strain to neomycin sulfate (NEO), ampicillin (AMP) and streptomycin (STR) reached 400 μg / mL or above, and the resistance to gentamicin (GEN) Resistance to 200 μg / mL, resistance to carbenicillin (CARB) and kanamycin (KAN) to 50 μg / mL, resistance to chloramphenicol (CM) to 20 μg / mL, resistance to tetracycline (TC) resistance is less than 5 μg / mL. This result is helpful for selecting appropriate antibiotics as a reference in follow-up studies.

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Abstract

The invention discloses a quorum sensing signal molecule DSF quenching strain and application of the quorum sensing signal molecule DSF (Diffusible Signal Factor) quenching strain. The research findsthat the Ralstonia pickettii has an obvious effect of degrading a quorum sensing signal molecule DSF, meanwhile, Ralstonia pickettii F20 capable of efficiently degrading the DSF is screened, and the strain is preserved in the Guangdong Microbiological Culture Collection Center on April 3, 2018 with the preservation number of GDMCC NO: 60347. The strain can rapidly efficiently degrade the DSF, so that a DSF-mediated quorum sensing system is interfered, the disease caused by the DSF is obviously reduced, and the function of the bio-control effect is achieved. The chemical control method is replaced, the potential safety hazard in the use of antibiotics is reduced, and new exploration and idea for the researches on the control technique of microbe diseases are provided.

Description

technical field [0001] The invention belongs to the technical field of plant disease biological control. More specifically, it relates to a quorum sensing signal molecule DSF quenching bacteria and its application. Background technique [0002] The quorum sensing system of the DSF family is a conserved cellular communication system widely found in Gram-negative bacteria. The quorum sensing system of the DSF family can be divided into three categories: the first category is based on the cruciferous plant pathogen Xanthomonas campestris ( Xanthomonas campestris pv. campestris , Xcc ) as a representative, this type of DSF quorum sensing system has confirmed the presence of Xanthomonas ( Xanthomonas sp.), Xylella fastidiosa (X ylella fastidiosa ), Lysobacterium enzymogenes ( Lysobacter enzymogenes ) and Stenotrophomonas maltophilia ( Stenotrophomonas maltophilia ); the second type of DSF family quorum sensing system is based on the opportunistic pathogen Burkholde...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/01
CPCA01N63/00C12N1/20C12N1/205C12R2001/01
Inventor 陈少华何杰华郭徨纯袁昊张玥黄少莉张炼辉
Owner SOUTH CHINA AGRI UNIV
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