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Pseudomonas fluorescens and its application in the control of tobacco black shank

A technology of Pseudomonas fluorescens and tobacco black shank, applied in the field of microbiology, can solve the problems of poor control effect of tobacco black shank, and achieve the effect of active and stable, simple operation method and high activity

Inactive Publication Date: 2020-01-14
陈德鑫 +7
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the above-mentioned problem of poor control effect of tobacco black shank in the prior art, the present invention provides a Pseudomonas fluorescens that has an antagonistic effect on tobacco black shank, and a method for preventing and treating tobacco black shank using the technology

Method used

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  • Pseudomonas fluorescens and its application in the control of tobacco black shank
  • Pseudomonas fluorescens and its application in the control of tobacco black shank
  • Pseudomonas fluorescens and its application in the control of tobacco black shank

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The screening of embodiment 1 Pseudomonas fluorescens

[0020] The screening method of Pseudomonas fluorescens of the present invention has the following steps:

[0021] 1. Isolation of soil bacteria

[0022] Take 10g of soil and add 90mL to take 10g of the collected soil sample, add it to a conical flask filled with 90mL of sterile water, place the conical flask on a shaker, shake at 180rpm for 20min, and make 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, and absorb 200 μL of each concentration on the NA plate On, spread evenly, repeat six times for each concentration. Then the coated NA plates were incubated in a constant temperature incubator at 28°C for 24h.

[0023] 2. Primary screening

[0024] Inoculate the bacteria cake of Geotrichum candidum (Geotrichum candidum) with a diameter of 9 mm in the center of a PDA plate...

Embodiment 2

[0028] The identification of embodiment 2 Pseudomonas fluorescens

[0029] Biochemical identification was performed on the bacteria obtained above, which showed that they were Pseudomonas fluorescens.

[0030] 1. Physiological and biochemical identification

[0031] Gram staining, starch hydrolysis, oxidase, nitric acid reduction, gelatin liquefaction, etc., the determination of physiological and biochemical characteristics refers to "Plant Pathogenic Bacteria Identification Experiment Guide"

[0032] The physiological and biochemical identification of the XF10 strain is shown in Table 1. The XF10 strain is a Gram-negative bacterium that can liquefy gelatin and reduce nitrate, but cannot hydrolyze starch, and the methyl red and V-P tests are negative. The strain was preliminarily identified as Pseudomonas. The results of the salt tolerance test were positive for 1% Nacl and negative for 4% Nacl.

[0033] Table 1 Physiological and biochemical identification of strain XF10

...

Embodiment 3

[0062] The Antagonism of Example 3 Bacteria Liquid to Tobacco Black Leg Disease

[0063] 1) Cultivation of tobacco black shank: in an aseptic operating table, a 0.5 cm diameter bacterial block was taken out with a puncher, inoculated in oat medium, and cultured in the dark at 28° C. for 3-5 days.

[0064] 2) Bacteria inoculation: Insert a 5mm-diameter black shank bacterial block in the center of the oat medium plate, inoculate 10 μL of XF10 strain bacterial solution on the left and right sides 24 mm from the center, repeat 3 times, culture in dark for 72 hours, and measure the size of the inhibition zone , to calculate its inhibition rate.

[0065] Inhibition rate (%) = (CK net growth amount - treatment net growth amount) / CK net growth amount × 100

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Abstract

The invention provides a strain of Pseudomonas fluorescens XF10, the preservation number of which is: CGMCC No.13703. The bacterial liquid, extracellular metabolites and volatile products of Pseudomonas fluorescens can effectively inhibit tobacco black shank. The Pseudomonas fluorescens screened by the present invention has high and stable activity, and its fermentation liquid, extracellular metabolites and volatile products can effectively kill tobacco black shank bacteria, adopts the root irrigation method, and the operation method is simple; The environment does not pose a hazard.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to Pseudomonas fluorescens which has antagonistic effect on tobacco black shank and application thereof. Background technique [0002] Tobacco black shank (Tobacco black shank) is a devastating soil-borne disease in tobacco production, and it is also the most serious root disease in tobacco production in the world. Tobacco black shank can harm the entire tobacco growth period, but mainly harms the tobacco plants in the field stage. The onset of the seedling stage often causes "damping-off" symptoms. First, black spots appear at the base of the stem, or develop from the bottom leaves and then extend to the stem along the petiole. Excessive humidity in the seedbed or rainy weather often cause the tobacco seedlings to die in pieces, and the diseased seedlings are covered with white hyphae; when the climate is dry, the diseased seedlings are brown and dry. The roots, stems, l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01N3/00C12R1/39
CPCA01N63/00A01N63/10C12N1/205C12R2001/39
Inventor 张瑞平陈德鑫杨艺炜刁朝强李斌王晓强王奇张立猛李茂业
Owner 陈德鑫
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