A dsf quorum sensing signal molecule quenching bacterium and its application in plant disease control

A quorum sensing signal and molecular technology, applied in applications, plant growth regulators, microorganism-based methods, etc., can solve the problem of rare DSF degrading bacteria, and reduce the problem of antibiotic abuse, pesticide residue pollution, black rot disease The degree of lightening, the effect of good adaptability

Active Publication Date: 2020-08-07
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, studies on the quenching mechanism of quorum-sensing signal molecules in the DSF family of Xanthomonas campestris have been carried out at home and abroad, but there are still few reports on the degrading microorganisms of DSF and / or DSF signal molecule analogs, and DSF-degrading bacteria are more as rare

Method used

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  • A dsf quorum sensing signal molecule quenching bacterium and its application in plant disease control
  • A dsf quorum sensing signal molecule quenching bacterium and its application in plant disease control
  • A dsf quorum sensing signal molecule quenching bacterium and its application in plant disease control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Isolation and identification of bacterial strain HN-2

[0044] 1. Isolation and screening of strains

[0045] (1) The soil was collected from the rice rhizosphere soil in the experimental field of South China Agricultural University in Guangzhou City, Guangdong Province, and the soil was yellowish brown.

[0046] (2) Isolation and purification of strains

[0047] Prepare MM medium, divide it into 250 mL Erlenmeyer flasks, fill each bottle with 50 mL of MM medium, sterilize it in a high-pressure sterilizer, and add DSF mother solution in the ultra-clean workbench after it cools down to make the culture medium The final concentration of DSF in the medium was 50 μm / L, and 5 g of soil sample was added to the medium at the same time. After culturing in a shaker at 30 °C and 200 rpm for 7 days, the 10% inoculum was transferred to the second batch of MM medium with a DSF concentration of 100 μm / L. After 7 days of culture under the same conditions, 10% of the inocu...

Embodiment 2

[0060] Example 2 Antibiotic susceptibility analysis of Copper greedy bacteria HN-2

[0061] In order to better study the biocontrol potential of the bacterial strain HN-2 obtained in Example 1, we studied the antibiotic sensitivity of the bacterial strain.

[0062] The result is as Figure 4 As shown, the strain was resistant to rifampicin (RIF) at 400 μg / mL or more, and to chloramphenicol (CM), tetracycline (TC), gentamicin (GEN), ampicillin (AMP) and Kanamycin (KAN) resistance is less than 10 μg / mL.

[0063] The results showed that strain HN-2 exhibited high resistance to rifampicin and low resistance to chloramphenicol, tetracycline, gentamycin, ampicillin, and kanamycin. It is beneficial to select appropriate antibiotics as a reference in follow-up research.

Embodiment 3

[0064] Embodiment 3 Determination of growth and DSF degradation relation curve of copper greedy bacteria HN-2

[0065] 1. Pick a single colony of strain HN-2 and inoculate it in LB medium to pre-culture to the logarithmic phase. After centrifuging the obtained bacterial solution at 4000 rpm for 5 min, discard the supernatant, and use 0.9% anhydrous Rinse and resuspend bacteria in normal saline, as a seed suspension, inoculate 50 mL of MM basal medium with an inoculum of 1-3%, and add DSF mother solution to make the final concentration 2mM, at 30°C, 200 rpm cultured and sampled at regular intervals. Collect samples at different time points and measure OD by spectrophotometer 600 The value represents the growth of strain HN-2, and the residual amount of DSF determined by HPLC represents the degradation of DSF by strain HN-2.

[0066] 2. The relationship between the growth of strain HN-2 and the degradation of DSF when DSF is used as the only carbon source is as follows Figur...

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Abstract

The invention discloses a DSF quorum sensing signal molecule quenching bacterium and its application in plant disease control. The present invention studies and finds copper greedy bacteria ( Cupriavidus sp.) can grow and reproduce with DSF as the sole carbon source, nitrogen source and energy source, and has a significant degradation effect on higher concentrations of DSF; at the same time, the present invention also provides a strain HN-copper greedy bacteria that efficiently degrades DSF 2. The strain was deposited in the Guangdong Microbial Culture Collection Center on August 13, 2018, with the preservation number GDMCC NO: 60432. Copper greedy bacteria HN‑2 has significant degradative activity on quorum sensing signal molecules in plant pathogens, and is environmentally friendly, and has great applications in the control of pathogenic plant pathogens mediated by DSF and / or DSF signal molecule analogs At the same time, the invention can reduce the problem of antibiotic abuse and pesticide residue pollution, reduce environmental pressure, provide a new idea for biological control of plant diseases, and has great application value and broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of biological control. More specifically, it relates to a DSF quorum sensing signal molecule quenching bacterium and its application in plant disease control. Background technique [0002] DSF (Diffusible Signal Factor) signal molecule is derived from Gram-negative pathogenic bacteria and is an unsaturated fatty acid substance. DSF signaling molecules are involved in the regulation of important biological functions such as microbial cell growth, biofilm formation, and production of pathogenic factors. Studies have found that the quorum sensing system mediated by DSF signaling molecules not only widely exists in Xanthomonas ( Xanthomonas ) bacteria, but also in Stenotrophomonas maltophilia ( Stenotrophomonas maltophilia ), Xylella fastidiosa ( Xylella fastidiosa ), Pseudomonas aeruginosa ( Pseudomonas aeruginosa ), Burkholderia ( Burkholderia ) and many marine bacteria. In addition, a variety o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P1/00C12R1/01
CPCA01N63/00A01N63/10C12N1/205C12R2001/01
Inventor 陈少华叶田林子秋范兴辉李绮婷罗青青张炼辉
Owner SOUTH CHINA AGRI UNIV
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