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Pseudomonas nitroreducens and application thereof in degradation of DSF

A technology of Pseudomonas and signal molecules, applied in the field of biological control, can solve problems such as the small number of degradable DSF strains, achieve significant biodegradation effect, good biocontrol effect, and reduce the problem of pesticide abuse.

Active Publication Date: 2018-01-26
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are many studies on the quorum sensing phenomenon of different plant pathogens and the biosynthesis mechanism of quorum sensing DSF signal molecules at home and abroad, but there are still few reports on the microbial degradation of DSF, and the number of strains that can degrade DSF is very small. Degradation of bacterial strains is mostly mentioned as a broad-spectrum degradation test, and its growth and degradation properties need to be further studied

Method used

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  • Pseudomonas nitroreducens and application thereof in degradation of DSF
  • Pseudomonas nitroreducens and application thereof in degradation of DSF
  • Pseudomonas nitroreducens and application thereof in degradation of DSF

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Acquisition and identification of Pseudomonas nitroreductor strain HS-18

[0047] 1. Isolation and screening of strain HS-18

[0048] (1) Soil sample collection: collected from long-term oil polluted soil as a microbial source.

[0049] Soil samples were collected on February 9, 2014 from the soil contaminated by food oil for a long time near South China Agricultural University in Guangzhou City, Guangdong Province. The soil from the surface to the depth of 5cm was sampled, bagged, and stored as a microbial source and brought back to the school for strains separate.

[0050] (2) Prepare culture medium: use DSF as the only carbon source, prepare LB medium and liquid MM inorganic salt medium and solid MM inorganic salt medium containing 0.5mMDSF without any other carbon source, use hydrochloric acid and sodium hydroxide Adjust the pH to 7.0, and sterilize at 121°C for 20 minutes.

[0051] Wherein, the inorganic composition and concentration of MM inorganic sa...

Embodiment 2

[0066] Example 2 Antibiotic susceptibility analysis of strain HS-18

[0067] In order to better study the biocontrol potential of the strain HS-18 obtained in Example 1, we conducted an in-depth study on the biological characteristics of the strain HS-18.

[0068] The sensitivity of strain HS-18 to different antibiotics was investigated experimentally, such as Figure 5 shown.

[0069] The experimental results show that the strain HS-18 has strong resistance to ampicillin (amp), chloramphenicol (cm), carbenicillin (carb), streptomycin (str), etc., and the resistance range reaches 200μg ·mL -1 Above; sensitive to gentamicin (gen), kanamycin (kan), etc., and the resistance range is less than 10 μg·mL -1 .

Embodiment 3

[0070] Example 3 Determination of Degradation Ability of Strain HS-18 to DSF

[0071] 1. Strain culture and sample collection: The strain HS-18 was streaked on LB solid medium plates and cultured at 30°C overnight. Pick a single colony, cultivate it in LB liquid medium overnight, use it as a seed solution, and then inoculate it with 1:100 inoculum in MM inorganic salt medium with 0.5mM DSF as the sole carbon source, at 30°C and 150rpm at a constant temperature Incubate in a shaker for 24h.

[0072] Samples were collected at different time points and tested as follows:

[0073] First, the absorbance value at wavelength 600nm was measured by an ultraviolet spectrophotometer (NANODROP) to indicate the growth of microorganisms; then ethyl acetate was added to another sample for extraction, the upper organic phase was taken, repeated twice, and the extracts were combined , after rotary evaporation to dryness, dilute the volume to 500 μL with methanol (HPLC pure) and put it in a h...

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Abstract

The invention discloses a pseudomonas nitroreducens and application thereof in degradation of DSF (diffusible signal factor) and / or DSF analogues. The pseudomonas nitroreducens disclosed by the invention is named as HS-18, and is preserved in China Center for Type Culture Collection on May 12, 2017 with a preservation number of CCTCC NO:M2017257. The strain has stable activity, can take DSF as thesole carbon source for growth, can completely degrade DSF within 24h, has a significant and rapid degradation effect on DSF, and has high application value in quorum quenching direction, black rot disease inhibition, and control of the harm of DSF mediation dependent pathogenic bacteria. The pseudomonas nitroreducens provided by the invention can reduce the pesticide abuse problem, and at the same time provides a new idea for biological control of the harm of phytopathogen.

Description

technical field [0001] The invention relates to the technical field of biological control, more specifically, to a nitroreducing pseudomonas (Pseudomonas nitroreducens) and its application in degrading quorum sensing signal molecule DSF and DSF signal analogues. Background technique [0002] The effective control of plant pathogenic bacteria has always been an important problem in agricultural production. For example, Xanthomonas bacteria are a class of Gram-negative plant pathogenic bacteria. They can infect more than 400 kinds of plants, including many important food crops. Crops and commercial crops, such as: rice, cotton, wheat, soybeans, citrus, bananas, etc., lead to a reduction in the production of these crops, causing major economic losses. Xanthomonas campestris (X. campestrispv. campestris, Xcc) contains at least 141 pathogenic species, which can infect all cruciferous plants and cause black rot. Black rot is an important agricultural disease, and there is no very...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P1/00C12R1/38
Inventor 张炼辉王惠杉陈少华范兴辉叶田
Owner SOUTH CHINA AGRI UNIV
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