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Myxococcus stipitatus preying on plant pathogenic bacteria and application thereof in biological control of bacterial diseases

A technology of plant pathogenic bacteria and pathogenic bacteria, applied in the field of applied microbiology, can solve problems such as differences in antibacterial effects, and achieve the effect of huge application potential

Active Publication Date: 2018-09-21
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are large differences in the antibacterial effects of different species of strains. For example, Bacillus shows different antibacterial properties in terms of resistance to plant pathogens, and related strains have also applied for patent protection (201380042989.6; 201410164779.0 and 201510666311.6, etc.)

Method used

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  • Myxococcus stipitatus preying on plant pathogenic bacteria and application thereof in biological control of bacterial diseases
  • Myxococcus stipitatus preying on plant pathogenic bacteria and application thereof in biological control of bacterial diseases
  • Myxococcus stipitatus preying on plant pathogenic bacteria and application thereof in biological control of bacterial diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1. Separation and purification of bacterial strains

[0032] 1.1 After the sample is collected, let it dry naturally at room temperature (to reduce the pollution of mold and miscellaneous bacteria).

[0033] Soil samples collected from Anhui and other places were air-dried naturally at room temperature.

[0034] 1.2 Induction of myxobacteria fruiting bodies

[0035] 1.2.1 Fecal particle induction plate method: Put about 20 grams of air-dried soil samples in a petri dish, half bury 3-4 sterilized rabbit feces on the surface of the soil samples, add 25 μg / ml ether sterilized put After soaking for 6 hours with Cycloheximide (25mg / ml) or Carbendazim (30mg / l), pour off the residual liquid, incubate at 30°C, and observe the formation of fruiting bodies on the feces ball after 48 hours.

[0036] 1.2.2 Yeast induction plate method: use WCX (cycloheximide 25mg / ml) medium as the basic medium, pour it into a sterile plate, draw a cross line with yeast on the surface of ...

Embodiment 2

[0047] Example 2. Identification of bacterial strains

[0048] 2.1 Morphological features

[0049] The isolated and screened myxobacteria BS were negative by Gram staining. Observation under the oil microscope by crystal violet staining revealed that the shape of the single bacterium was rod-shaped, without flagella, and both ends were transparent. Layer mucus layer; find that this bacterial strain has colony effect in the process of growth by light microscope 10 * observation, and growth late stage bacterium colony gathers together to form fruiting body ( figure 2 ).

[0050] 2.2 Identification

[0051] The 16S rDNA gene sequence of the strain was obtained by amplifying the 16S rDNA of the strain by PCR, and logged in NCBI (www.ncbi.nlm.nih.govPblastP) to obtain the 16S rDNA sequence of a typical strain similar to the experimental strain. The obtained sequencing results were compared with the known sequences in the database using BLAST software in the GenBank database. Th...

Embodiment 3

[0052] Example 3. Research on the predation effect of Myxococcus stipitatus BS on plant pathogenic bacteria 3.1 Research on the predation effect of strain BS on various plant pathogenic bacteria.

[0053] Ea, Dickeya fangzhongdai, Dickeya chrysanthemi, Dickeya solani, Pcc, RS were inoculated in LB liquid medium (tryptone 1%, NaCl 1%, yeast extract 0.5%), cultivated overnight at 30°C, centrifuged at 8000rpm for 3min, collected the bacteria, and used Resuspend in sterile water to OD 600 for 10. Drop it into TPM (10mM Tris-HCl [pH7.6], 1mM KH2PO4 [pH 7.6], 8mM MgSO 4 )) Blow dry on a solid plate for later use. Inoculate strain BS (CCTCCM2017496) into CYE liquid medium (Casitione 1%, Yaest extract 0.5% MgSO 4 ·7H 2 (0.1%pH 7.2-7.6), cultivated at 30°C for 2 days, centrifuged at 8000rpm for 3min, collected the thallus, resuspended the thallus with sterile water, and the thalline concentration was 10 8 cells / l, hanging drop on the air-dried pathogenic bacteria lawn, culture at ...

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Abstract

The invention discloses myxococcus stipitatus preying on plant pathogenic bacteria and application thereof in biological control of bacterial diseases. Slime bacteria BS of antagonistic plant pathogenic bacteria is provided, and the deposit number is CCTCCNO. M2017496. The myxococcus stipitatus BS inhibits the growth of various plant pathogenic bacteria such as cabbage soft rot pathogen, calla soft rot pathogen, tomato bacterial wilt pathogen, potato black smut and pear fire blight by predation, and shows slime bacteria BS broad spectrum resistance to pathogenic bacteria. Pot experiments showthat the slime bacteria BS can effectively inhibit the infection of Pectobacterium carotovorum subsp. carotovorum on potatoes and calla, reduce the incidence of soft rot, and can be used for the biological control of the plant diseases caused by plant pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of applied microbiology, and relates to a petiole myxococcus that preys on plant pathogenic bacteria and its application in biological control of bacterial diseases. Background technique [0002] Plant disease is one of the important restrictive factors for high-quality and high-yield crops. It is estimated that the average disease loss of major crops in the world accounts for nearly 14% of the total output, and the annual direct economic loss is as high as hundreds of billions of dollars. In order to deal with the harm caused by plant diseases, chemical pesticide control and crop breeding have become important means at present. However, the environmental pollution caused by chemical pesticides and the limitation of long breeding cycle make environment-friendly bio-control methods become the focus of research. focus. Microbial biological control technology has the characteristics of high efficiency, no pollution, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04A01N63/00A01N63/02A01P1/00C05G3/00C12R1/01
CPCA01N63/00A01N63/10C05G3/00C12N1/20C12P1/04C12N1/205C12R2001/01
Inventor 崔中利李周坤王婷罗雪黄彦
Owner NANJING AGRICULTURAL UNIVERSITY
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