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Application of phage trp574 gene in reducing the resistance of Ralstonia solanacearum to phage

A phage-resistant, bacterial wilt technology, applied in phage, virus/phage, microorganism-based methods, etc., can solve problems such as ineffective control, and achieve the effect of improving the use and control effects

Active Publication Date: 2022-05-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the control methods for bacterial wilt of crops mainly include chemical control, biological control, planting of disease-resistant varieties, crop rotation, etc. Although these measures have played a certain role in controlling bacterial wilt, they still cannot effectively control the disease.

Method used

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  • Application of phage trp574 gene in reducing the resistance of Ralstonia solanacearum to phage
  • Application of phage trp574 gene in reducing the resistance of Ralstonia solanacearum to phage
  • Application of phage trp574 gene in reducing the resistance of Ralstonia solanacearum to phage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

O2.32μL, the total system is 10μL. The PCR program is: 94°C 5min; 94°C 30s, 55°C 30s,

72°C for 40s, 30 cycles; 72°C for 5min, 16°C. Liquid culture the strain containing the target band, extract the plasmid and use

BamHI and Hind III were digested for verification, and the recombinant plasmid was handed over to Invitrogen for sequencing and identification. obtained recombination

The plasmid was named pBBR‑orf30.

Table 2 orf30-F / orf30-R and MCS-F / MCS-R primer sequence

[0044]

2, result

(1) according to phage genome sequence design primer and PCR amplification to obtain the target fragment of about 688bp (Fig. 1A),

The target fragment is sequenced, and it is found that the orf30 sequence is 555bp, and its nucleotide sequence is as shown in SEQ ID NO: 1, and its

Embodiment 2

[0050] The preparation of the R. solanacearum Tb15 electric shock competent state and the electric shock transformation method refer to the method of Lavie et al. (2002).

[0052] (2) Add 1 mL of 10% glycerol to resuspend, centrifuge at 6000 rpm at 4° C. for 5 min, and discard the supernatant.

[0054] (4) 100 μL of 10% glycerol was added to resuspend to obtain R. solanacearum competence.

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Abstract

The invention discloses the application of the phage trp574 gene in reducing the resistance of R. solanacearum to phages. The research of the present invention shows that the phage trp574 gene can eliminate the resistance of R. solanacearum to phages and make the resistance sensitive; The source gene trp574 is transferred to R. solanacearum strains, and the transgenic R. solanacearum strains with reduced resistance to phages can be obtained, thereby improving the use and control effect of phages, and used for the prevention and treatment of R. solanacearum.

Description

Application of bacteriophage trp574 gene in reducing phage resistance of R. solanacearum technical field The present invention relates to the technical field of plant disease control, more particularly, relate to a kind of bacteriophage trp574 gene in reducing Application of R. solanacearum to phage resistance. Background technique Ralstonia solanacearum (Ralstonia solanacearum, referred to as Ralstonia solanacearum) is a gram-negative plant disease The original fungus, widely distributed in tropical, subtropical and temperate regions, can infect more than 450 kinds of plants in more than 50 families, and is the main cause of bacterial wilt in crops. of pathogenic bacteria. The fungus invades from the roots of plants, first colonizes the intercellular spaces of the root cortex, and then colonizes the ducts and adjacent tissues. It proliferates rapidly and spreads widely, resulting in blockage and destruction of the vascular system, which eventually leads to the with...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/74C12N1/21C12R1/01
CPCC12N15/74C07K14/005C12N2795/00022
Inventor 刘琼光胡蓉花余成鹏钟敏
Owner SOUTH CHINA AGRI UNIV
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