Pseudomonas. chlororaphis subsp. Aurantiaca Pa40 and application thereof

A technology of orange subspecies and green needle false single, applied in the direction of application, chemicals for biological control, biocides, etc., to achieve the effect of high research and application value

Inactive Publication Date: 2009-08-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the use of Pseudomonas chloropinus subsp

Method used

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  • Pseudomonas. chlororaphis subsp. Aurantiaca Pa40 and application thereof
  • Pseudomonas. chlororaphis subsp. Aurantiaca Pa40 and application thereof
  • Pseudomonas. chlororaphis subsp. Aurantiaca Pa40 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The identification of embodiment 1 Pa40

[0028] Based on the morphological characteristics, physiological and biochemical characteristics, 16S rDNA sequence, atpD gene sequence, recA gene sequence, carA gene sequence and BIOLOG carbon source metabolic profile of Pseudomonas, it was identified as Pseudomonas chloropinus subsp. Pseudomonas. chlorophis subsp. aurantiaca. The specific identification results are as follows:

[0029] 1. Morphological characteristics of bacteria

[0030] Gram staining was negative, electron microscope observation, bacteria oblong.

[0031] 2. Physiological and biochemical characteristics

[0032] The physiological and biochemical characteristics of Pseudomonas.chlorophis subsp.Aurantiaca Pa40 CGMCCNo.2764 are shown in Table 1: Gram-negative bacteria, strict aerobic, nitrate reduction, starch hydrolysis, esculin utilization, Negative α-galactase reaction, oxidase, arginine double hydrolysis, gelatin hydrolysis, lecithinase, lipase, glucose...

Embodiment 2

[0040] Embodiment 2 antibacterial spectrum analysis

[0041] Plate inhibition test method for pathogenic fungi. Take a 5mm-diameter target pathogenic fungus and inoculate it in the center of the PDA plate. Pseudomonas and the target fungus are inoculated at 2.5cm from both sides of the target pathogenic fungus at an angle of 180°. After culturing at 25°C for 4 to 6 days, measure the colony diameter of the pathogenic fungus . Plates inoculated with target fungi but not pseudomonas were used as controls. The growth inhibition rate was calculated according to the following formula:

[0042] Growth inhibition rate=(C-T) / C×100%

[0043] C: Control fungal growth diameter

[0044] T: diameter of fungal growth after inoculation with pseudomonas

[0045] Plate inhibition detection method for pathogenic bacteria. The inhibition of Pseudomonas to pathogenic bacteria was detected by double-layer culture method. Use King`B (peptone 20g, glycerin 10ml, K 2 HPO 4 1.5g, MgSO 4 ·7H ...

Embodiment 3

[0055] Example 3 Detection of biocontrol-related traits

[0056] Protease detection: use skim milk plate (tryptone 5g, yeast extract 2.5g, glucose 1g, 7% skim milk 250ml, agar 15g, add 1000ml of water) to detect protease. Inoculate Pa40 into the center of the plate and incubate at 28°C for 72 hours. There will be a transparent circle around the colony, such as image 3 As shown, Pseudomonas chlororaphis subsp.aurantiaca Pa40 CGMCCNo.2764 produces protease.

[0057] HCN detection: HCN test paper preparation, 10mg copper (II) ethyl acetoacetate (copper(II) ethyl acetoacetate) and 10mg 4,4'-methylbis(xylaniline) [4,4'-methylenebis-(N,N-dimethylaniline) ] dissolved in 4ml of chloroform, cut Whatman filter paper strips, soaked in the prepared solution, after the chloroform volatilized completely, HCN detection test paper was prepared. Puncture and inoculate pseudomonas in a centrifuge tube containing 1ml of PDA medium, suspend the HCN detection test paper in the centrifuge tube, ...

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Abstract

The invention provides a Pseudomonas.chlororaphis subsp.Aurantiaca Pa40, preservation number of which is CGMCCNo.2764. The strain generates HCN, prolease and siderophore relative to antibacterium and has plate board inhibition effect on a majority of tested plant pathogenic fungi and a part of tested plant pathogenic bacteria. Control effect of the biocontrol strain Pa40 on rhizoctonia solani is detected by taking wheat as an indicator plant on the plate, in the greenhouses and in the fields; a result shows that the control effects of the Pa40 on the rhizoctonia solani on the plate bed, in the greenhouse and in the fields are respectively 53.97%, 68.10% and 72.46%; wherein the control effects on the rhizoctonia solani in the greenhouses and fields are higher than that of validamycin.

Description

technical field [0001] The invention relates to a new microbial strain and its application, in particular to a Pseudomonas chloropinus subsp. orange strain and its application in biological disease control. Background technique [0002] Wheat sheath blight (Rhizoctonia cerealis), also known as blight blight and sharp eye spot, is one of the worldwide diseases with wide distribution. Mainly occurs in temperate wheat regions of the world. It was reported abroad as early as 1934. Although this disease has occurred earlier in our country, the harm is relatively light. Since the mid-to-late 1970s, with the replacement of wheat varieties and the popularization and application of high-yield cultivation measures (such as early sowing, irrigation, high fertilizer, etc.), the disease has generally occurred in various winter wheat regions in my country, and has a tendency to increase year by year. The disease area expanded from south to north, posing a greater threat to the high and...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/38
Inventor 郭岩彬邬娜吴文良孟凡乔赵桂慎
Owner CHINA AGRI UNIV
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