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Myxococcus xanthus preying on ralstonia solanacearum and application of myxococcus xanthus in biological prevention and control of tomato bacterial wilt

A R. solanacearum and yellow technology, applied in the field of microorganisms, can solve the problems of unstable control effect, poor control effect in the later stage, poor economic traits of disease-resistant varieties, etc.

Active Publication Date: 2021-10-22
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complex field environment, environmental conditions have a greater impact on its colonization and the activity of secondary metabolites. When it is actually used in agricultural production, problems such as unstable control effect and poor control effect in the later stage often occur.
Planting disease-resistant varieties is the main control measure in production at present, but the economic traits of existing disease-resistant varieties are often poor. Therefore, developing more efficient control methods for tomato bacterial wilt is an important research direction

Method used

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  • Myxococcus xanthus preying on ralstonia solanacearum and application of myxococcus xanthus in biological prevention and control of tomato bacterial wilt
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  • Myxococcus xanthus preying on ralstonia solanacearum and application of myxococcus xanthus in biological prevention and control of tomato bacterial wilt

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: the separation and purification of bacterial strain ( figure 1 )

[0024] 1.1 After the samples were collected, they were taken back to the laboratory and dried naturally at room temperature.

[0025] Soil samples were collected from tomato bacterial wilt disease-infected fields on the farm of South China Agricultural University and air-dried naturally at room temperature.

[0026] 1.2 Induction of fruiting bodies of myxobacteria

[0027] 1.2.1 Ralstonia solanacearum (prey) plate induction: water agar plate (CaCl 2 2H 2 (2.01g / L, 3-morpholinepropanesulfonic acid 2.093g / L, agar 15g / L, pH 7.2) is the base separation medium, prepares the aseptic water agar solid plate (adding final concentration before pouring plate is 25mg / mL Cycloheximide to inhibit the growth of enzyme bacteria), on the surface of the culture medium, draw a field grid with R. solanacearum cultured in advance, put a small amount of soil samples in the four square boxes of the field grid...

Embodiment 2

[0032] Example 2. Identification of bacterial strains

[0033] 2.1 Morphological identification

[0034] The myxobacteria R31 isolated and screened from the soil of the bacterial wilt disease area was negative for Gram staining, and observed under an optical microscope through crystal violet staining, it was found that the shape of the single cell was rod-shaped, with transparent ends and no flagella It is found that there is a layer of mucus layer around the R31 thalline by transmission electron microscope observation; it is found that the R31 bacterial strain has group behavior in the growth process by stereoscope 1 observation, and the bacterial colony in the growth stage gathers together to form a fruiting body structure visible to the naked eye ( figure 2 ).

[0035] 2.2 Molecular biological identification

[0036] By extracting the DNA of strain R31, the 16S rDNA gene sequence of R31 was amplified by using the bacterial 16S rDNA specific primers 27F and 1492R and Taq ...

Embodiment 3

[0037] Example 3. The predation ability and functional evaluation of Myxococcus xanthus R31 strain on Ralstonia solanacearum

[0038] 3.1 The predation effect of Myxococcus xanthus R31 on multiple strains of Ralstonia solanacearum with different pathogenicity

[0039] Scrape an appropriate amount of the activated Myxococcus xanthus R31 strain on the VY / 2 plate and inoculate it in CTT liquid medium, culture it in a shaker at 28°C and 180rpm for 2 days, then centrifuge at 8000rpm for 5min to collect the bacteria. TPM buffer (CTT liquid medium without casein peptone) was resuspended to prepare Myxococcus xanthus R31 suspension. Pick a single colony of activated R. solanacearum in TM liquid medium (10 g / L bacteriological peptone, 1 g / L acid hydrolyzed peptone, 5 g / L glucose, pH 7.0), and culture in a shaker at 30°C and 180 rpm , cultivated for 36-48h, and when the cells grew to the logarithmic growth phase, centrifuged at 8000rpm for 5min to collect the bacteria, resuspended with...

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Abstract

The invention discloses myxococcus xanthus preying on ralstonia solanacearum and application of the myxococcus xanthus in biological prevention and control of tomato bacterial wilt. The invention discloses Myxococcus xanthus R31, and the preservation number of the Myxococcus xanthus R31 is GDMCC No: 61842. According to the invention, plant pathogen ralstonia solanacearum is innovatively adopted as a predatory bacterium, a myxobacteria strain R31 is successfully separated and screened from a collected soil sample by using a predatory bacterium induction method, the myxobacteria strain R31 is identified as myxobacteria xanthus, the strain can prey on multiple strains of ralstonia solanacearum with different pathogenicity in the growth process and pathogenic bacteria are split to provide nutrients for self growth. The myxobacteria preys on ralstonia solanacearum, and physiological and biochemical characteristics and physiological differentiation of strains are not distinguished, so that broad-spectrum resistance of the myxobacteria to ralstonia solanacearum is shown. On the basis of predation experiments and potting experiments of Zhongshu No.4 tomatoes, experiments show that the myxobacteria Myxococcusx anthus R31 has huge application potential in the aspect of biological prevention and control of tomato bacterial wilt.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a Myxococcus xanthus that preys on Ralstonia solanacearum of tomato and its application in biological control of Ralstonia solanacearum of tomato. Background technique [0002] Bacteria wilt (Bacteria wilt) is an important disease in tomato production. It is widely distributed in the world and often breaks out in high temperature and high humidity environments (such as in southern provinces and cities in my country). Bacterial wilt has an acute onset and spreads quickly. When it is severe, it will cause the death of plants in patches, severely reduce production, or even fail to harvest. Tomato bacterial wilt, known as the "cancer" of tomato, has become one of the important constraints that limit the quality and high yield of tomato worldwide. According to reports, the annual yield loss caused by bacterial wilt is about 15%-95%. The pathogen of tomato bacterial...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/20A01P1/00C12R1/01
CPCC12N1/20A01N63/20
Inventor 董红红朱红惠姚青徐欣高瑞祥李安章
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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