Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria

A technology for Pseudomonas and pathogenic bacteria, applied in the field of microorganisms, can solve the problems of polluted environment, pesticide residues, long degradation cycle and so on

Inactive Publication Date: 2016-05-04
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the short production cycle of vegetables, traditional chemical pesticides have problems such as environmental pollution, long degradation cyc

Method used

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  • Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria
  • Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria
  • Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Isolation and preliminary screening of antagonistic bacteria

[0018] The method of Fang Zhongda (1979) was used to isolate the growth-promoting bacteria in the rhizosphere. Take 10 g of the collected tobacco rhizosphere soil, add it into a conical flask filled with 90 mL of sterile water, place the conical flask on a shaker, and vibrate at 180 rpm for 20 min to prepare a soil suspension. Let the prepared soil suspension stand for 5 minutes, take the supernatant, and use the 10-fold serial dilution method to successively dilute to 10 -2 、10 -3 、10 -4 and 10 -5 , pipette 100 μL of each concentration on the NBY plate, spread evenly, and repeat each concentration three times. Then the coated NBY plates were incubated in a constant temperature incubator at 28°C for 24h. When bacterial colonies appeared on the NBY plate, the bacterial suspension of Ralstonia solanacearum (2.0×10 8 wxya -1 ) evenly sprayed on the NBY plates that have grown colonies, and cult...

Embodiment 2

[0020] Embodiment 2: Determination of antagonistic bacteria inhibition spectrum

[0021] Pick a single colony of antagonistic bacteria XQ10 and culture it in 5ml NBY liquid medium at 28°C with shaking at 200rpm for 16-24h. Collect the bacteria by centrifugation, adjust the bacterial concentration to OD with sterile water 420 =0.3 (about 2.0×10 8 wxya -1 ). Pipette 10 μL of the above bacterial suspension to the center of an NBY plate with a diameter of 90 mm. After the surface of the medium is fully absorbed, transfer the plate to a 28°C biochemical incubator for 72 hours. Spray OD 420 =0.3 for the indicator bacteria suspension, count the bacteriostasis after 24 hours. The results of the bacterial antagonism experiment showed that the antagonist XQ10 was effective against the common bacterial pathogens R. The bacteria (P.carotovora) all had significant antagonistic activity, and the size of the inhibition zone against R. solanacearum was 25.67±0.27mm.

Embodiment 3

[0022] Embodiment 3: Physiological and biochemical identification of antagonistic bacteria

[0023] Prepare a culture box, add 5mL sterile water to the box to establish a humid environment, and put the reagent strips into the box. Pick a single colony, resuspend it with sterile saline to prepare a bacterial suspension, and adjust the concentration of the bacterial suspension to OD 420= 0.3. Use a sterile syringe to draw the physiological saline bacterial solution into the test tube from NO3 to PNPG (the cup part does not need to be added). Add mineral oil to the GLU, ADH and URE test tubes until a convex crescent is formed. Open the ampoule of AUX medium, add 200μL (4-8 drops) of the remaining normal saline bacterial solution, mix well and avoid the formation of air bubbles. The test tube from GLU to PAC should be filled with AUX bacterial solution, and the surface of the tube should be flat or slightly convex, but not concave or protruding crescent. Cover and incubate at ...

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Abstract

The invention belongs to the field of microorganisms, and relates to separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria. The strain XQ10 is separated from tobacco rhizosphere. An API 20 NE parenteral gram-negative bacillus identification result shows that XQ10 belongs to pseudomonas; 16S rRNA and multilocus sequence typing analysis based on gyrB, rpoB and rpoD show that the strain XQ10 belongs to pseudomonas but has a large difference from reported strains in pseudomonas, the strain XQ10 is regarded as a new strain which is not reported yet, and thus the strain is named Pseudomonas shandongensis XQ10. An antagonistic experiment shows that the strain has a significant inhibition effect on various plant pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of microorganisms, and relates to the isolation and application of a strain of Pseudomonas which has a significant antagonistic effect on plant pathogenic bacteria, in particular to an antagonism against Ralstonia solanacearum (Ralstonia solanacearum) and soft rot of Chinese cabbage in crops such as tobacco and peanuts (Pectobacterium carotovora) and tomato bacterial canker bacteria (Clavibactermichiganensis) and other plant pathogenic bacteria Pseudomonas isolation, identification and application. Background technique [0002] Tobacco bacterial wilt is a typical systemic vascular disease caused by Ralstonia solanacearum, which mainly damages the vascular tissues of tobacco roots, stems and leaves, and is the main disease of tobacco in tropical and subtropical regions One, it generally occurs in the Yangtze River Basin of my country and the tobacco-growing areas to the south. In recent years, the occurrence and hazard...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/38
CPCA01N63/00C12N1/205C12R2001/38
Inventor 李向东王晓强李亦晴
Owner SHANDONG AGRICULTURAL UNIVERSITY
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