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Pseudomonas donghuensis 22G5 and application thereof in prevention and treatment of crop verticillium wilt

A technology for pseudomonas and verticillium wilt, applied in the field of plant disease biocontrol, can solve problems such as limited strains and products, and achieve the effects of reducing the incidence rate, inhibiting infection, and reducing the occurrence of verticillium wilt

Inactive Publication Date: 2020-01-14
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current biological control strains and products for cotton Verticillium wilt are very limited, and only a few products containing active microbial components such as active Bacillus, Trichoderma or Pseudomonas fluorescens can be used to prevent and control Verticillium wilt

Method used

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  • Pseudomonas donghuensis 22G5 and application thereof in prevention and treatment of crop verticillium wilt
  • Pseudomonas donghuensis 22G5 and application thereof in prevention and treatment of crop verticillium wilt
  • Pseudomonas donghuensis 22G5 and application thereof in prevention and treatment of crop verticillium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Isolation and screening of embodiment 1 Pseudomonas 22G5

[0031] The experimental procedures for large-scale screening of Verticillium wilt antagonistic strains are attached figure 1 as shown in a:

[0032] 1. Sample Collection

[0033] Samples were collected from Anhui, Zhejiang, Jiangsu, Xinjiang, Jilin, Heilongjiang, Hunan, Shanghai and other places. Weeds, corn, cotton, Soiled root samples of various plants such as soybeans. The root samples with soil were isolated by dilution coating plate method. Select soil, root system and other samples respectively, place them in 50ml collection tubes, add 45ml water to mix and shake, generally carry out 4-5 times of shaking, absorb 800μl of the mixed solution, and use ddH 2 O Perform 10 on a 96-well plate 1 -10 5 gradient dilution. Select 10 μl solution of appropriate concentration to spread on LB medium, generally choose 10 μl 3 or 10 4 The diluted concentration of the plate was spread on the plate, and the plate wa...

Embodiment 2

[0036] Example 2 Identification and genome sequencing of Pseudomonas fluorescens 22G5

[0037] 1. Morphological identification

[0038] Strain 22G5 is Gram-negative, non-spore-forming, and provides motility with single or multiple flagella. After being cultured on LB medium for 24 hours, large colonies can be formed. The colonies do not produce pigment, and the surface is raised, smooth, viscous and easy to provoke.

[0039] 2. Molecular Identification

[0040] In order to clarify the strain classification of the bacterial strain 22G5 obtained in Example 1, we analyzed the 16S ribose DNA (rDNA) sequence of the bacterial strain 22G5 for the molecular biology identification of bacteria. The homology of the bacterial model strain Pseudomonas donghuensis HYS (T) strain reaches the highest 99%, indicating that the strain belongs to Pseudomonas donghuensis species. Pseudomonas is a class of Gram-negative bacteria containing multiple species. Due to their diversity in morphologica...

Embodiment 3

[0045] The mensuration of embodiment 3 Pseudomonas 22G5 antibacterial activity

[0046] In order to better study the biocontrol potential of the bacterial strain 22G5 of the present invention, we studied the bacteriostatic spectrum of the bacterium.

[0047] The specific operation is as follows:

[0048] Zone of inhibition test

[0049] In order to explore whether 22G5 has broad-spectrum resistance to multiple strong pathogenic strains of Verticillium dahliae, use the same method as in Example 1 for strain screening, and replace the pathogenic bacteria V15QY1 with the same amount of culture conditions and methods. Multiple strong pathogenic strains of V07df2, V08df1 and V991, the rest of the steps remain unchanged. Experimental results found that strain 22G5 also had a strong antibacterial effect on Verticillium dahliae V07df2, V08df1 and V991 ( image 3 ).

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Abstract

The invention discloses a pseudomonas donghuensis 22G5 and an application of the pseudomonas donghuensis 22G5 in prevention and treatment of crop verticillium wilt. The pseudomonas donghuensis 22G5 ispreserved in the China General Microbiological Culture Collection Center on July 08, 2019, and a preservation number of the pseudomonas donghuensis 22G5 is CGMCC No. 18084. The pseudomonas donghuensis 22G5 has no pathogenicity to plants, has a remarkable inhibition effect on cotton verticillium wilt caused by Verticillium dahliae, can remarkably reduce the occurrence of cotton verticillium wilt,and has a very good biological control effect. The genome studies show, the pseudomonas donghuensis 22G5 has a gene cluster for synthesizing gene cluster of siderophore 7-hydroxycycloheptyltrienol ketone, and the compound can be used as an iron ion chelating agent to inhibit the iron element necessary for the growth of verticillium dahliae, so that the growth of pathogenic bacteria is inhibited. The strain can be used for developing biopesticides for crop verticillium wilt, and provides a new thought and a new method for biological prevention and treatment of crop verticillium wilt.

Description

technical field [0001] The invention belongs to the technical field of plant disease biocontrol. More specifically, it relates to a strain of Pseudomonas donghuensis 22G5 and its application in the control of crop verticillium wilt. Background technique [0002] Cotton is an important economic crop in my country, providing renewable raw materials for the textile industry. Cotton verticillium wilt caused by Verticillium dahliae Kleb. is a devastating disease. The spread of the disease not only significantly reduces the cotton fiber quality, but also seriously affects the economic yield of cotton. Verticillium wilt caused by Verticillium dahliae causes direct economic losses in the billions of dollars worldwide each year. In 1982, cotton verticillium wilt had spread to nearly 130,000 hectares of agricultural soil in my country. By 1993, cotton verticillium wilt had become the main obstacle to high and stable cotton production in my country, and the affected area reached 2.6...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00A01G7/06C12R1/39
CPCA01G7/06A01N63/00C12N1/205C12R2001/39
Inventor 关雪莹陶晓园张海琳
Owner ZHEJIANG UNIV
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