A Nitroreductor Pseudomonas and its Application in Degrading Quorum Sensing Signaling Molecule DSF

A quorum sensing signal, Pseudomonas technology, applied in the direction of microorganism-based methods, bacteria, chemicals for biological control, etc., can solve the problem of small number of degradable DSF strains, achieve significant biodegradation effect, reduce The problem of pesticide abuse, the effect of good biocontrol effect

Active Publication Date: 2020-08-07
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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  • A Nitroreductor Pseudomonas and its Application in Degrading Quorum Sensing Signaling Molecule DSF
  • A Nitroreductor Pseudomonas and its Application in Degrading Quorum Sensing Signaling Molecule DSF
  • A Nitroreductor Pseudomonas and its Application in Degrading Quorum Sensing Signaling Molecule 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] Antibiotic susceptibility analysis of embodiment 2 bacterial strain HS-18

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

[0068] The sensitivity of the strain HS-18 to different antibiotics was experimentally studied, 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 200 μg ·mL -1 above; more sensitive to gentamicin (gen), kanamycin (kan), etc., and the resistance range is less than 10 μg·mL -1 .

Embodiment 3

[0070] Degradability determination of embodiment 3 bacterial strain HS-18 to DSF

[0071] 1. Strain cultivation and sample collection: Streak the strain HS-18 on the LB solid medium plate and cultivate overnight at 30°C. Pick a single colony, culture it with LB liquid medium overnight, as the seed solution, and inoculate it in the MM inorganic salt medium with 0.5mM DSF as the only carbon source at an inoculum size of 1:100, keep the temperature at 30°C and 150rpm Incubate in a shaker for 24 hours.

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

[0073] First use a UV spectrophotometer (NANODROP) to measure the absorbance value at a wavelength of 600nm to indicate the growth of microorganisms; then add ethyl acetate to another sample for extraction, take the upper organic phase, repeat twice, and combine the extracts After rotary evaporation to dryness, dilute to 500 μL with methanol (HPLC pure) and place in a high-performance liquid chromatograp...

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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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Patent Type & Authority Patents(China)
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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