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Pseudomonas chlororaphis and application thereof in prevention and treatment of plant bacterial soft rot

A technology based on Pseudomonas chloropinus and bacteria, which is applied in the direction of bacteria, applications, plant growth regulators, etc., can solve the problem of unknown chemical structure of VFM quorum sensing signal

Active Publication Date: 2022-05-13
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, due to the unknown chemical structure of the VFM quorum sensing signal, it has become one of the barriers to the development of Dickeya soft rot fungus QQ technology

Method used

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  • Pseudomonas chlororaphis and application thereof in prevention and treatment of plant bacterial soft rot
  • Pseudomonas chlororaphis and application thereof in prevention and treatment of plant bacterial soft rot
  • Pseudomonas chlororaphis and application thereof in prevention and treatment of plant bacterial soft rot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation and screening of bacterial strain L5

[0030] 1. Strain isolation

[0031] Sample collection: Field soil samples were collected in rice fields in Xiaogan, Hubei, stored in bags, and brought back to the laboratory for strain isolation.

[0032] Isolation of bacterial strains: Soil samples were isolated by dilution coating plate method. Specifically, after fully mixing the collected field soil samples, 5.0 g was weighed and placed in a sterilized centrifuge tube filled with 15 mL of fresh LB liquid medium, shaken at 28°C and 200 rpm for 15 minutes, and then allowed to stand until the soil Particles sink to the bottom of the centrifuge tube, the supernatant is diluted with sterile water, and 100 μL of 10 -5 、10 -6 、10 -7 The diluted solution was spread on an LB plate, placed upside down in a constant temperature incubator at 28°C for 24 hours, and then the obtained single colony was streaked and purified. Pick a single colony on the purified plate...

Embodiment 2

[0035] Example 2: Flow cytometry verification of the quenching effect of strain L5 on VFM

[0036] The VR2 reporter strain was activated by streaking with LB+Kan solid medium, and a single colony was picked and placed in LB+Kan liquid medium, and cultured on a shaker at 28°C and 200rpm until the bacterial liquid OD 600 = 1.0, the reporter strain VR2 and bacterial strain L5 (quenched bacteria) were inoculated and mixed in equal amounts, and the reporter strain VR2 was only inoculated as a control, and the fluorescence intensity MFI of the sample was measured every four hours. From figure 2 It can be seen that when the strain is cultivated for 4-8 hours, the strain L5 can significantly quench the VFM signal produced by VR2, and when the strain is cultivated for 12 hours, the quenching gene of VR2 can degrade the VFM signal in the system. And without adding strain L5, the VFM signal value that the reporter system responded to decreased sharply compared with the original.

Embodiment 3

[0037] Example 3: Identification of strain L5

[0038] 1. Morphological identification

[0039] Strain L5 is Gram-negative; colonies on LB nutrient medium are orange-yellow, round, raised, smooth, viscous, easy to pick up, and neatly edged.

[0040] 2. Molecular identification

[0041] In order to clarify the taxonomic status of the strain L5 obtained in Example 1, we identified the strain L5 using the method of multilocus sequence analysis (MLSA) combined with phylogenetic analysis (Phylogenetic analysis).

[0042] 16S rDNA sequence analysis is a molecular biological identification method for bacterial identification. By analyzing the 16S rDNA sequence (the nucleotide sequence is shown in SEQ ID NO.1), combined with the conserved housekeeping gene rpoB (the nucleotide sequence is shown in SEQ ID NO. 2) and rpoD sequence (the nucleotide sequence is shown in SEQ ID NO.3), using MEGA 6.0 software to construct a phylogenetic tree using the Neighbor-joining method, and comprehen...

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Abstract

The invention discloses pseudomonas chlororaphis and application of the pseudomonas chlororaphis in prevention and treatment of plant bacterial soft rot. The preservation number of the pseudomonas chlororaphis L5 is GDMCCNO.62059. The pseudomonas chlororaphis L5 has a very good quenching effect on a VFM quorum sensing signal generated by Dickeya soft rot fungi, can remarkably reduce the occurrence of bacterial soft rot of various crops, and has a good prevention and treatment effect in a pot experiment; moreover, the pseudomonas chlororaphis L5 has no pathogenicity, can be used for developing a biocontrol agent for the crop bacterial soft rot, and provides a new thought and a new material for biologically preventing and treating the crop bacterial soft rot.

Description

technical field [0001] The invention belongs to the technical field of plant disease biocontrol, and in particular relates to a strain of Pseudomonas chloropinus and its application in preventing and treating plant bacterial soft rot. Background technique [0002] Quorum sensing (QS) is a cell-to-cell communication process that enables bacteria to collectively change behavior in response to changes in the cell density and environment of the surrounding microbial community. In the past ten years, the quorum sensing system between bacteria has become a research hotspot in the field of microbiology. It not only reveals the regulatory network that affects the important life processes of microorganisms, but also brings new hope for the research and treatment of microbial infectious diseases. Quorum quenching (QQ) refers to blocking the signal exchange between bacteria by interfering with or destroying the quorum sensing system without producing drug resistance and without affecti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/27A01P1/00C12R1/38
CPCC12N1/20A01N63/27Y02A40/10
Inventor 周佳暖刘帆胡明王思张炼辉
Owner SOUTH CHINA AGRI UNIV
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