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Acinetobacter lactucae and application thereof to degradation of quorum-sensing signal molecule DSF

A quorum sensing signal and signal molecule technology, applied in applications, bacteria, animal repellants, etc., can solve the problems of the small number of DSF degrading strains and the inability to meet the biological control of diseases, reduce the pollution of pesticide residues, and greatly promote the application. Potential, effectiveness in reducing the problem of antibiotic misuse

Active Publication Date: 2018-04-27
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the research on the quenching mechanism of quorum sensing signal molecules of Xanthomonas campestris DSF family has been carried out at home and abroad, but there are still few reports on the microbial degradation of DSF, and the number of DSF-degrading strains screened is very small, which is far from satisfying. The need for biological disease control

Method used

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  • Acinetobacter lactucae and application thereof to degradation of quorum-sensing signal molecule DSF
  • Acinetobacter lactucae and application thereof to degradation of quorum-sensing signal molecule DSF
  • Acinetobacter lactucae and application thereof to degradation of quorum-sensing signal molecule DSF

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Experimental program
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Effect test

Embodiment 1

[0043] The acquisition and identification of embodiment 1 Acinetobacter strain QL-1

[0044] 1. Isolation and screening of strain QL-1

[0045] (1) Soil sample collection: The rice rhizosphere soil collected from the test field was used as the microbial source.

[0046] The soil samples were collected from the rice rhizosphere soil in the experimental field of South China Agricultural University in Guangzhou City, Guangdong Province on July 16, 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.

[0047] (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. After culturing on a shaker at 30 ° C and 200 rpm for 7 days,...

Embodiment 2

[0075] Antibiotic susceptibility analysis of embodiment 2 bacterial strain QL-1

[0076] In order to better study the biocontrol potential of the bacterial strain QL-1 obtained in Example 1, we studied the antibiotic sensitivity of the bacterial strain QL-1. Such as Figure 5 As shown, the strain is resistant to chloramphenicol (CM) up to 200 μg / mL or more, to ampicillin (AMP) up to 50 μg / mL, to rifampicin (RIF) and carbenicillin (CARB) Resistance reaches 20μg·mL -1 , the resistance to streptomycin (STR), kanamycin (KAN), gentamicin (GEN), tetracycline (TET) is less than 10 μg / mL. This result is helpful for selecting appropriate antibiotics as a reference in follow-up studies.

Embodiment 3

[0077] The determination of the growth and degradation DSF relation curve of embodiment 3 bacterial strain QL-1

[0078] 1. Pick a single colony of strain QL-1 and inoculate it in LB medium to pre-culture to the logarithmic phase. After the obtained bacterial solution is centrifuged at 4000rpm for 5 minutes, the supernatant is discarded, and the bacterial cells are washed with 0.9% sterile saline and weighed. Suspended, as a seed suspension, inoculated into 50mL MSM basal medium at an inoculum size of 1:100, and added DSF mother solution to make the final concentration 2mM, cultured at 30°C, 200rpm for 24h, and took samples regularly. Collect samples at different time points and measure OD by spectrophotometer 600 The value represents the growth situation of bacterial strain QL-1, and the residual amount of DSF measured by HPLC represents the degradation situation of bacterial strain QL-1 to DSF.

[0079] 2. The results of HPLC detection are as follows: Figure 6 Shown (wher...

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Abstract

The invention discloses Acinetobacter lactucae and an application thereof to degradation of a quorum-sensing signal molecule DSF (diffusible signal factor). The invention provides a bacterial strain of Acinetobacter lactucae QL-1, the bacterial strain is preserved in a China Center for Type Culture Collection on September 11th, 2017, and the preservation number is CCTCC NO: M2017487. The bacterialstrain can employ the quorum-sensing signal molecule DSF as a sole carbon source for growth, and 2mM of DSF signal molecules can be completely degraded in 15 hours with high degradation activity andstable degradation performance. The bacterial strain QL-1 has substantial biocontrol effects for black rot caused by Xanthomonas campestris XC1, and the bacterial strain also has characteristics of fast growth speed, simple cultivation method, good adaptation capability without easy variation, and great application potential for promotion of control of pathogens which are pathogenic depending on DSF; and at the same time, the product can reduce the problem of abuse of pesticides, and provides a new thinking for biological control of plant diseases.

Description

technical field [0001] The invention belongs to the technical field of plant disease biological control. More specifically, it relates to an Acinetobacter lactucae and its application in degrading quorum sensing signal molecule DSF. Background technique [0002] Many microbial signaling molecules have been discovered and isolated one after another. These signaling molecules regulate the expression of specific microbial genes, especially a variety of virulence factors, and coordinate group behavior, that is, quorum sensing. DSF (Diffusible Signal Factor) signaling molecule is a kind of unsaturated fatty acid derived from Gram-negative pathogenic bacteria. Like the quorum sensing system that is more researched on AHLs, it also regulates the formation of biofilm, cell growth and pathogenic factors. production and other important biological functions. Studies have found that quorum sensing mediated by DSF signaling molecules is highly conserved in many plant pathogenic bacteri...

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 陈少华叶田张炼辉范兴辉王惠杉阳芳冯芷萱单雯艳
Owner SOUTH CHINA AGRI UNIV
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