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DSF quorum sensing signal molecule quenching bacterium and application thereof in plant disease prevention and treatment

A quorum sensing signal and molecular technology, applied in the fields of application, plant growth regulator, botanical equipment and methods, etc., can solve the problem of rare DSF degrading bacteria, reduce the problem of antibiotic abuse and pesticide residue pollution, and achieve high degradation activity , the effect of reducing environmental pressure

Active Publication Date: 2018-12-25
SOUTH CHINA AGRI UNIV
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  • Description
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  • 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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  • DSF quorum sensing signal molecule quenching bacterium and application thereof in plant disease prevention and treatment
  • DSF quorum sensing signal molecule quenching bacterium and application thereof in plant disease prevention and treatment
  • DSF quorum sensing signal molecule quenching bacterium and application thereof in plant disease prevention and treatment

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 1-3% inoculum into 50 mL MM basal medium, and add DSF mother solution to make the final concentration 2 mM, at 30°C, 200 rpm cultured under the conditions and sampled regularly. 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 Fig...

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Abstract

The invention discloses a DSF quorum sensing signal molecule quenching bacterium and application thereof in plant disease prevention and treatment. Researches of the invention discover that Cupriavidus sp. can grow and propagate by using DSF as the only carbon source, nitrogen source and energy and has an evident degradation effect on high-concentration DSF. The Cupriavidus sp. strain HN-2 which is capable of efficiently degrading DSF is preserved in Guangdong Microbial Culture Collection Center on August 13th, 2018, and the preservation number of the strain is GDMCC No. 60432. The Cupriavidussp. strain HN-2 has evident degradation activity to quorum sensing signal molecules in phytopathogens, is environmentally friendly, has huge application potential in the prevention and treatment of DSF and / or DSF signal molecule analogue mediated phytopathogen, can reduce the problems of antibiotics abuse and pesticide residue pollution, can relieve environment pressure, provides a new thought for biological plant disease prevention and treatment, and has a huge application value and promising application prospect.

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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IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P1/00C12R1/01
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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