A pathogenic strain causing patchouli bacterial wilt and its application

A technology of patchouli and bacterial wilt, which is applied to the field of pathogenic strains that cause patchouli bacterial wilt, and achieves a high affinity effect

Active Publication Date: 2022-05-10
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether Kosakonia can cause patchouli bacterial wilt has not been reported

Method used

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  • A pathogenic strain causing patchouli bacterial wilt and its application
  • A pathogenic strain causing patchouli bacterial wilt and its application
  • A pathogenic strain causing patchouli bacterial wilt and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Isolation and purification of Patchouli solanacearum and pathogenicity assay

[0037] (1) Separation and purification: collect patchouli plants with typical symptoms of bacterial wilt disease from the field, soak the diseased plants in tissue, inoculate the infusion solution on a TTC plate, culture at 30°C for 48 hours, and pick out bacteria that are related to bacterial wilt Single colonies with similar colony morphology were purified and preserved.

[0038]Among them, the culture medium of TTC plate is acid hydrolyzed casein 1.0g / L, glucose 5.0g / L, peptone 10.0g / L, agar powder 10g / L and TTC 0.05g / L, pH7.2.

[0039] (2) Observation of colony morphology: inoculate the purified and preserved strains on a TTC plate by the three-line method, culture at 30° C. for 24 hours, and observe the bacterial colony morphology. The colonies are smooth circular bulges, with a red center and a white bacterial circle, with obvious fluidity, such as figure 1 shown.

[0040] ...

Embodiment 216

[0041] Example 2 16s rDNA sequence identification

[0042] Genomic DNA of strain Pa82 was used as a template to perform PCR amplification of 16s rDNA gene. The primers were: 27F: 5′-AGAGTTTGATCCTGGCTCAG-3′, 1541R: 5′-AAGGAGGTGATCCAGCCGCA-3′. Use 25 μL system for PCR reaction: 1 μL each of 27F / 1541R (10 μM), 1.5 μL of genomic DNA, ddH 2 O 9.0 μL, Taq enzyme 12.5 μL; reaction parameters: pre-denaturation at 94°C for 2 min; denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 100 s, 35 cycles; extension at 72°C for 10 min. PCR samples were subjected to agarose gel electrophoresis ( Figure 4 A), and sequence the samples. Compare the sequence results of the 16s rDNA gene on NCBI, select strains with high similarity and other plant pathogenic bacterial strains, and use the Neighbor-joining method in MEGA5.1 software to construct the phylogenetic tree of 16s rDNA, as shown in Figure 5 shown. The results showed that the strain Pa82 was clustered with ...

Embodiment 3

[0044] Example 3 Identification of rpoB gene sequence

[0045] Genomic DNA of strain Pa82 was used as a template to perform PCR amplification of rpoB gene, and the primers were: rpoB F: 5′-ATGGTTTACTCCCTATACCGAGA-3′, rpoB R: 5′-TTACTCGTCTTTCCAGCTCGAT-3′. PCR reaction uses 25 μL system: rpoB F / R primer (10 μM) 1 μL each, genomic DNA 1.5 μL, ddH 2 O 9.0 μL, Taq enzyme 12.5 μL; reaction parameters: pre-denaturation at 94°C for 2 min; denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 100 s, 35 cycles; extension at 72°C for 10 min. PCR samples were subjected to agarose gel electrophoresis ( Figure 4 B), and sequence the samples. Compare the sequence results of the rpoB gene on NCBI, select strains with high similarity and other plant pathogenic bacterial strains, and use the Neighbor-joining method in the MEGA5.1 software to construct a phylogenetic tree of the rpoB gene, as shown in Figure 6 shown. The results showed that the strain Pa82 was cl...

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Abstract

The invention discloses a pathogenic strain causing patchouli bacterial wilt and application thereof, belonging to the technical field of detection of plant pathogenic bacteria. The pathogenic strain is Kosakonia sp.Pa82, the preservation number: GDMCC No: 61047, which is a new pathogenic strain of patchouli bacterial wilt. The pathogenic strain has strong pathogenicity to Patchouli, which provides research materials for the study of R. Screening, breeding of disease-resistant varieties, and research and development of plant vaccines against bacterial wilt are of great significance, and provide research ideas for the prevention and control of bacterial wilt in other plants.

Description

technical field [0001] The invention belongs to the technical field of detection of plant pathogenic bacteria, in particular to a pathogenic strain causing patchouli bacterial wilt and application thereof. Background technique [0002] Patchouli bacterial wilt is a vascular disease caused by bacteria. Pathogens invade from root wounds of plants, colonize and multiply in vascular bundles, cause difficulties in water transportation of plants, cause stems and leaves to wilt, and eventually wither and die. Pathogenic bacteria can overwinter in soil and diseased tissues, and the disease is more serious in continuous cropping land. Bacteria wilt can occur throughout the growth period of plants, and is most prevalent in high temperature and rainy seasons. Once a plant is infected, it spreads rapidly, resulting in the death of a large area of ​​plants. Bacteria wilt is a devastating disease in patchouli production. The isolation and identification of pathogenic bacteria is very ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/18A01N63/20A01P1/00C12R1/01
CPCC12Q1/18A01N63/20C12R2001/01C12N1/205
Inventor 贺红李巧张宇瑶张泳
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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