DSF quorum sensing signal degrading bacteria, and applications thereof in plant disease control

A technology of quorum sensing signal and degrading bacteria, which is applied in the fields of application, plant growth regulator, botanical equipment and methods, etc., can solve problems such as interference, achieve stable degradation performance, reduce the problem of antibiotic abuse, and reduce the problem of pesticide residue pollution Effect

Active Publication Date: 2018-04-20
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In recent years, screening microorganisms from the environment or cloning degrading enzymes to degrade AHLs (N-Acyl homoserinelactones) signal molecules, interfering with the quorum sensing system using AHLs as signal molecules, and preventing and treating AHLs-mediated pathogenic bacteria have been the most in-depth research, but screening has There are still few reports on DSF-degrading strains with biocontrol potential

Method used

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  • DSF quorum sensing signal degrading bacteria, and applications thereof in plant disease control
  • DSF quorum sensing signal degrading bacteria, and applications thereof in plant disease control
  • DSF quorum sensing signal degrading bacteria, and applications thereof in plant disease control

Examples

Experimental program
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Effect test

Embodiment 1

[0045] The acquisition and identification of embodiment 1 Burkholderia bacterial strain HN-8

[0046] 1. Isolation and screening of strain HN-8

[0047] (1) Soil sample collection: The rice rhizosphere soil collected from the test field was used as the microbial source.

[0048] The soil samples were collected from the rice rhizosphere soil in the experimental field of South China Agricultural University in Guangzhou, Guangdong Province on June 23, 2016. The soil from the surface layer to the depth of 5 cm was sampled, bagged, and preserved as a microbial source for strain isolation.

[0049] (2) Enrichment culture of bacterial strains: prepare MSM medium, put 50mL of MSM medium into a 250mL Erlenmeyer flask for sterilization, add DSF mother liquor (methanol is solvent) under aseptic conditions after cooling, to make the final mass of DSF The concentration was 50 μM, and 5 g of soil samples were added at the same time, and after 7 days of incubation at 30 ° C and 200 rpm on a...

Embodiment 2

[0068] Antibiotic susceptibility analysis of embodiment 2 bacterial strain HN-8

[0069] In order to better study the biocontrol potential of the bacterial strain HN-8 obtained in Example 1, we studied the antibiotic sensitivity of the bacterial strain HN-8.

[0070] The result is as Figure 4 As shown, the resistance of the strain to ampicillin (AMP), carbenicillin (CARB), streptomycin (STR) reached 400 μg / mL or above, the resistance to tetracycline (TET) reached 200 μg / mL, and the resistance to Kanna The resistance to gentamicin (KAN) reaches 50 μg / mL, and the resistance to gentamicin (GEN) and neomycin sulfate (NS) reaches 10 μg·mL -1 , The resistance to rifampicin (RIF) and chloramphenicol (CM) is less than 10 μg / mL.

[0071] Antibiotic susceptibility results show that strain HN-8 has good antibiotic resistance, especially resistance to ampicillin, carbenicillin, and streptomycin. This result is not only conducive to the selection of appropriate antibiotics in subsequent...

Embodiment 3

[0072] Example 3 The hydrolysis circle experiment of bacterial strain HN-8 degrading DSF

[0073] In this example, the solid MSM medium plate with DSF as the sole carbon source was used to measure the hydrolysis circle generated by strain HN-8 degrading DSF.

[0074] 1. Prepare MSM solid medium, add a certain amount of DSF mother liquor as the only carbon source, so that the final concentration of DSF is 5mM. Pick a single colony of strain HN-8 and inoculate it in LB medium to pre-culture to the logarithmic phase. After centrifuging the obtained bacterial solution at 4000rpm for 5min, discard the supernatant, wash and resuspend the bacterial cells with 0.9% sterile normal saline, As a seed suspension. Take 0.5 μL of the seed suspension and place it on a solid MSM plate with 5 mM DSF as the sole carbon source, place the medium plate at a constant temperature of 30°C, and observe the hydrolysis circle after 72 hours. In order to ensure the reliability and accuracy of the exper...

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Abstract

The invention discloses a DSF quorum sensing signal degrading bacteria, and applications thereof in plant disease control, and more specifically provides Burkholderia anthina bacterial strain HN-8 preserved at the Guangdong Culture Collection Center, on 24th, November, 2017, and the preservation number is GDMCCNO.60289. It is found in study that Burkholderia anthina, especially the bacterial strain HN-8 possesses remarkable and rapid degrading effect on quorum sensing signal molecule DSF, the degradation performance is stable, quorum sensing communication can be destroyed via degradation of DSF, and great application potential is achieved in prevention and treatment of black rotten caused by xanthomonas campestris, and pathogenic bacteria based on DSF. The Burkholderia anthina bacterial strain HN-8 also possesses relatively high drug resistance on ampicillin, carbenicillin, and streptomycin, so that pesticide or antibiotic abuse and drug resistance are avoided, and novel methods are provided for prevention of plant diseases.

Description

technical field [0001] The invention belongs to the technical field of plant disease biocontrol. More specifically, it relates to a DSF quorum sensing signal degrading bacterium and its application in plant disease control. Background technique [0002] Quorum sensing (QS) refers to the phenomenon that microorganisms rely on a certain cell density to communicate between cells and coordinate group behavior. This coordinated process includes the synthesis and secretion of signal molecules, the perception of signal molecules and signal transduction, the activation of target gene expression, and finally a series of biological activities. DSF (Diffusible Signal Factor) signal molecule is a new type of quorum sensing signal synthesized and secreted by Xanthomonas, and its chemical structure is cis-11-methyl-2-dodecene. Successively, DSF and its structural analogs have been found in many microorganisms, such as Stenotrophomonas maltophilia, Xylella fastidiosa, Burkholderia cenoce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P1/00C12R1/01
CPCA01N63/00A01N63/10C12N1/205C12R2001/01
Inventor 陈少华冯芷萱张炼辉范兴辉王惠杉叶田阳芳
Owner SOUTH CHINA AGRI UNIV
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