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Method for detecting pathogenicity of kiwifruit bacterial canker

A canker bacteria and detection method technology, which is applied in the field of virulence detection of kiwifruit bacterial canker, can solve the problems of poor health, detection, and small amount of bacterial liquid, and achieves fast onset time, clear symptoms, and convenient sources. Effect

Inactive Publication Date: 2018-10-16
INST OF PLANT PROTECTION FAAS
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  • Abstract
  • Description
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Problems solved by technology

At present, the pathogenicity detection methods of kiwifruit bacterial canker are mostly used in the newborn kiwifruit detached leaves needle-spraying inoculation method and epidermis micro-bacteria injection inoculation method, but the following problems exist in this method: The amount of bacterial liquid is small, and the kiwifruit leaves under moisturizing conditions are prone to chlorosis 2 days after in vitro, and their health is far less than that of living plant leaves, which has a significant impact on the authenticity of the pathogenicity of the bacteria. Bacterial method, due to the inconvenient and inconsistent collection of materials, the onset time and inoculation effect are unstable, therefore, it is seldom used for the determination and evaluation of pathogenicity among different strains
[0004] Actinidia is a deciduous plant. The peak period of the disease is usually from the dormancy period to the budding stage of the plant, and the new leaves have not grown yet. The isolated strains cannot be tested for pathogenicity on the leaves of kiwifruit in time.

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  • Method for detecting pathogenicity of kiwifruit bacterial canker
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  • Method for detecting pathogenicity of kiwifruit bacterial canker

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preparation example Construction

[0034] Among the present invention, the preparation method of described kiwifruit bacterial canker bacteria suspension preferably comprises the steps:

[0035]A. Streak the bacterial strain of kiwifruit bacterial canker sore on the flat plate of peptone yeast powder medium, cultivate it in the dark for 72~96h under the condition of 25~28° C., and obtain the amplified kiwifruit bacterial canker sore;

[0036] B. Mix the kiwifruit bacterial canker bacteria after the expanded culture of the step A with sterile water to obtain a kiwifruit bacterial canker bacteria suspension.

[0037] In the present invention, the kiwifruit bacterial canker strain is preferably streaked on a plate of peptone yeast powder medium, and cultured in the dark at 25-28° C. for 72-96 hours to obtain the kiwifruit bacterial canker after expanded culture. In the present invention, the bacterial strain of kiwifruit bacterial canker is preferably activated before streaking on the plate. The activation method...

Embodiment 1

[0042] In the Fujian region, conventional diseased tissue isolation methods and kiwifruit bacterial canker specific molecular detection methods (refer to literature [Young Jin Koh, Ill Sup Nou. DNA Markers for Identification of Pseudomonas syringae pv. actinidiae. Mol. Cells, 2002, 13(2):309-314] method for specific molecular detection), single colonies obtained colonies showed pale milky white, strain numbers were Psafj01, Psafj02, Psafj03, Psafj04, Psafj05, Psafj06, Psafj07, Psafj08 kiwifruit bacterial ulcer Germ strains.

[0043] Among them, the conventional diseased tissue separation method: cut out kiwi fruit branch disease-like diseased tissue pieces with a size of about 1 to 2 cm, wash the diseased tissue pieces with sterile water 2 to 3 times, and disinfect with 75% ethanol for 30s to 60s. Mercury disinfection for 2 to 3 minutes, and then washed with sterile water for 2 to 3 times. Remove 3 to 5 pieces of sterilized diseased tissues, add 0.5 to 1 mL of sterile water, ...

Embodiment 2

[0046] After the kiwifruit bacterial canker bacterial strain obtained in Example 1 was activated with beef extract peptone culture solution, the activated kiwifruit bacterial canker bacterial strain Psafj01, Psafj02, Psafj03, Psafj04, Psafj05, Psafj06, Psafj07, Psafj08 and non-pathogenic bacteria NPsafj01 was streaked on the peptone yeast powder medium, and cultured in the dark at 25°C for 96 hours to obtain kiwifruit bacterial canker bacteria Psafj01, Psafj02, Psafj03, Psafj04, Psafj05, Psafj06, Psafj07, Psafj08 and non-pathogenic bacteria NPsafj01 after expansion culture , the kiwifruit bacterial canker bacterium after the expanded culture obtained is mixed with sterile water, so that the concentration of the kiwifruit bacterial canker bacterium suspension is 6×10 8 CFU / mL.

[0047] Puncture a small hole on the leaf of the tomato plant along the left and right sides of the vein, inoculate the suspension of Actinidia bacterial canker bacteria in the small hole, and obtain the...

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Abstract

The invention provides a method for detecting pathogenicity of kiwifruit bacterial canker, belonging to the technical field of crop pathogen detection. According to the method, bacterial suspension isused as an inoculum and is inoculated on the tomato leaves; obvious symptoms can be caused on the tomato leaves on the third day after the artificial inoculation; the symptoms are similar to those ofhost leaf spots of the kiwi fruit; the pathogenicity of the pathogen is determined by utilizing the characteristic that the symptoms of the tomato leaves are similar to those of kiwi fruit leaves. The method provided by the invention can meet the requirements of rapid detection of the pathogenicity of newly isolated pathogens and pathogens with different storage time at any time, and is better inapplicability.

Description

technical field [0001] The invention belongs to the technical field of crop pathogen detection, and in particular relates to a method for detecting the pathogenicity of kiwifruit bacterial canker. Background technique [0002] Kiwifruit bacterial canker (Kiwifruit bacterial canker) is a devastating bacterial disease in kiwifruit production, and is now listed as a national forest plant quarantine object. In recent years, with the rapid expansion of kiwifruit cultivation area, the spread of kiwifruit bacterial canker has gradually increased. The disease was discovered successively in Hunan, Anhui, Shaanxi, Sichuan, Chongqing and other places. During the epidemic years, the whole garden was on the verge of destruction and caused huge economic losses. Kiwifruit canker disease is caused by the infection of Pseudomonas syringae pv. It mainly harms the trunk, main branches and side branches. After the new leaves grow from April to May, the pathogen can transfer and infect the lea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
CPCC12Q1/02
Inventor 何玉仙杨秀娟代玉立兰成忠卢学松
Owner INST OF PLANT PROTECTION FAAS
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