A high-purity non-pathogenic Ralstonia solanacearum strain

A non-pathogenic technology for R. solanacearum, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of different adsorption capacity, non-pathogenicity, and different charges, and achieve stable biological properties , the effect of high purity

Active Publication Date: 2019-01-18
福建省农业科学院资源环境与土壤肥料研究所
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High purity and stable biological properties of the strain are important conditions for it to be an excellent strain of bacterial vaccines; it is difficult to separate and purify Ralstia solanacearum with traditional bacterial isolation techniques to obtain high-purity strains
R. solanacearum cell surface components with different pathogenicity are different, and their charges are also different, and their adsorption capacity with ion exchange resins is different. However, the following related reports have not been seen so far: Utilizing bacterial chromatography system can achieve Ralstonia solanacearum Purification and separation of R. solanacearum strains to obtain high-purity non-pathogenic R. solanacearum strains

Method used

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  • A high-purity non-pathogenic Ralstonia solanacearum strain
  • A high-purity non-pathogenic Ralstonia solanacearum strain
  • A high-purity non-pathogenic Ralstonia solanacearum strain

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Experimental program
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Effect test

Embodiment 1

[0014] The isolation of embodiment 1 bacterial strain FJAT-1957

[0015] (1) Weigh 5 g of tomato stems from the healthy plants of Lvhe Chengsheng Co., Ltd. in Jiaodang Village, Fuding City, Fujian Province, and rinse them with water, and cut the tomato stems after washing. Then, under aseptic conditions, the small pieces of tissue were treated with 75% alcohol for 30 seconds, sterilized with 10% sodium hypochlorite for 8 minutes, and rinsed with sterile water for 3 times; Obtain tissue fluid, and then draw 1mL of tissue fluid for gradient dilution, and the selected dilution is 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 and 10 -7 ;

[0016] (2) Spread 200 μL of the diluted solution obtained in step (1) on a TTC plate, and then place the TTC plate at 30° C. for 2 days;

[0017] (3) Streak-inoculate the strain obtained in step (2) on the TTC medium again, and culture the TTC medium at 30° C. for 48 h, and name the strain obtained from the culture as strain FJAT-1957.

Embodiment 2

[0018] Identification of embodiment 2 bacterial strain FJAT-1957:

[0019] Initial identification:

[0020] The colonies of the obtained bacterial strain FJAT-1957 were placed under a Leica M165FC advanced motorized fluorescent stereo microscope for microscopic observation. The observation results showed that the colony morphology of the bacterial strain FJAT-1957 was relatively dry on the surface, without fluidity, and dark red in the middle. And the strain with narrow white border, therefore, it can be preliminarily identified that the strain FJAT-1957 belongs to the genus Ralstia solanacearum.

[0021] Further identification:

[0022] Activation of strain FJAT-1957: Streak the obtained strain FJAT-1957 on the TTC medium with an inoculation loop, and place the TTC medium in a constant temperature incubator for 48 hours at a temperature of 30°C; the strain FJAT-1957 was obtained single colony;

[0023] Preparation of seed solution: Inoculate a single colony of the obtained...

Embodiment 3

[0032] Pathogenicity test of embodiment 3 bacterial strain FJAT-1957

[0033] Set up an experimental group, a negative control group and a positive control group. In the experimental group, the strain FJAT-1957 was activated on a TTC plate, inoculated in SPA liquid medium, and cultured at 170r / min and 30°C for 48h. diluted to 10 8 cfu / mL, then inoculate the roots of the diluted bacterial solution on tomato potted seedlings at the age of 5-6 (one tomato seedling is planted in each pot), the inoculation volume is 80 mL / plant, and a total of 30 tomato seedlings are inoculated; while negative The control group replaced the bacterial liquid dilution of bacterial strain FJAT-aRS01 with clear water; the positive control group replaced the bacterial liquid dilution of bacterial strain FJAT-1957 with the bacterial liquid dilution of the existing recognized R. solanacearum standard strain GMI1000; The treated tomato seedlings in each group were placed in a light incubator for cultivati...

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Abstract

The invention provides a high-purity avirulent Ralstonia solanacearum strain FJAT-aRS01(Ralstonia solanacearum) which is collected with the collection number of CGMCC No.10693 in CGMCC (China General Microbiological Culture Collection Center) located in No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing on April 7, 2015. The strain FJAT-aRS01 is high in purity, stable in biological trait and capable of effectively preventing and controlling crop bacterial wilt and provides possibility for preventing crop bacterial wilt through effectively researching and developing a bacterial wilt plant vaccine.

Description

【Technical field】 [0001] The invention belongs to the field of biology, and in particular relates to a high-purity non-pathogenic strain of Ralstia solanacearum. 【Background technique】 [0002] Crop bacterial wilt is a devastating soil-borne disease caused by Ralstonia solanacearum, which is difficult to control. The use of apathogenic R. solanacearum to develop plant vaccines against bacterial wilt has a good application prospect in preventing crop bacterial wilt. Inoculate the weak pathogenic strain - non-pathogenic Ralstia solanacearum at the seedling stage of the plant. After invasion and colonization, it will compete with the pathogenic bacteria for nutrients and sites in the plant, and build the micro-ecological balance in the plant. , Inducing plants to develop disease resistance, thereby preventing the spread of pathogenic bacteria and inhibiting the occurrence of diseases. [0003] The pathogenicity of R. solanacearum strains is severely differentiated in the natu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 刘波郑雪芳朱育菁唐建阳车建美
Owner 福建省农业科学院资源环境与土壤肥料研究所
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