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Pseudomonas protegens XY2F4 and application thereof in control of crop verticillium wilt

A Pseudomonas fluorescens, XY2F4 technology, applied in the direction of application, microbial-based methods, chemicals for biological control, etc., can solve the problems of limited strains and products, and achieve the effect of reducing the incidence rate and inhibiting infection

Active Publication Date: 2020-01-03
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 protegens XY2F4 and application thereof in control of crop verticillium wilt
  • Pseudomonas protegens XY2F4 and application thereof in control of crop verticillium wilt
  • Pseudomonas protegens XY2F4 and application thereof in control of crop verticillium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and Screening of Pseudomonas XY2F4

[0030] 1. Sample collection and strain cultivation: In the Yangtze River Basin, Xinjiang cotton production area, Hainan cotton southern breeding base, Northeast agricultural production area and other places near the soil, water body and plant root system, collected plant root samples with soil, bagged Save and bring back to the laboratory for strain isolation. The root samples with soil were isolated by dilution coating plate method. Select soil, root system and other samples respectively and place them in 50ml collection tubes, add 50ml water to mix and shake, generally carry out 4-5 times of shaking, absorb 800μl of 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 was incubated upside...

Embodiment 2

[0032] Example 2 Identification and genome sequencing of Pseudomonas fluorescens XY2F4

[0033] 1. Morphological identification

[0034] Strain XY2F4 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 can produce pigments, and the surface is raised, smooth, viscous and easy to provoke.

[0035] 2. Molecular identification

[0036] In order to clarify the bacterial species classification of the strain XY2F4 obtained in Example 1, we analyzed the 16S ribose DNA (rDNA) sequence of the strain XY2F4 for molecular biological identification of the bacteria. The results showed that the homology of the 16SrDNA sequence of XY2F4 with Pseudomonas protegens CHA0 and Pseudomonas protegens CHA0 and Pseudomonas saponiphila DSM9751 strains was up to 99%, indicating that the strains belonged to the large category of Pseudomonas. Pseudomonas is a class of Gram-...

Embodiment 3

[0044] The mensuration of embodiment 3 Pseudomonas XY2F4 antibacterial activity

[0045] In order to better study the biocontrol potential of the bacterial strain XY2F4 of the present invention, we have studied the bacteriostatic spectrum of the bacterium. The specific operation is as follows:

[0046] XY2F4 has the ability to inhibit the growth of other pathogenic fungi. Using the same method as in Example 1 for strain screening, the pathogenic bacteria V15QY1 was replaced with equal amounts of V07DF2, V08DF2, V08T2, and V16DF4 with the same culture conditions and methods, and the rest of the steps were unchanged. The experimental results found that the strain XY2F4 had a strong antibacterial effect on Verticillium dahliae V07DF2, V08DF2, V08T2, and V16DF4 ( image 3 a).

[0047] In addition, XY2F4 can also inhibit the growth of Verticillium dahliae. Verticillium dahliae for testing is a common and highly pathogenic strain of Verticillium dahliae V991. The single clones ...

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Abstract

The invention discloses Pseudomonas protegens XY2F4 and application thereof in control of crop verticillium wilt. The Pseudomonas protegens XY2F4 was collected in the China General Microbiological Culture Collection Center (CGMCC) of the China Committee for Culture Collection of Microorganisms (CCCCM) on June 24, 2019, with the collection number of CGMCC No.18017. The pseudomonas protegens XY2F4 has no virulence for plants, has a significant inhibition function on cotton verticillium wilt caused by verticillium dahliae, can remarkably relieve occurrence of cotton verticillium wilt and has goodbiological control effects. Genome research shows that the Pseudomonas protegens XY2F4 has a series of biological control related compound synthesis gene clusters, can be used for developing biological pesticides for crop verticillium wilt diseases, and provides a new thought and a new method for biological control of the 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 protegens XY2F4 and its application in controlling 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 2.666 m...

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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