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Application of cycloserine and beneficial bacteria in preventing and controlling tomato soil-borne ralstonia solanacearum

A technology of cycloserine and beneficial bacteria, applied in the field of microorganisms, can solve problems such as limited nutrients

Active Publication Date: 2020-09-01
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the nutrients in the soil are limited, and when the beneficial bacteria are used to prevent and control tomato soil-borne diseases, it is often difficult for the beneficial bacteria to proliferate in large quantities in the soil and exert their due control effects.

Method used

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  • Application of cycloserine and beneficial bacteria in preventing and controlling tomato soil-borne ralstonia solanacearum
  • Application of cycloserine and beneficial bacteria in preventing and controlling tomato soil-borne ralstonia solanacearum
  • Application of cycloserine and beneficial bacteria in preventing and controlling tomato soil-borne ralstonia solanacearum

Examples

Experimental program
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Effect test

Embodiment 1

[0020] The Indoor Effect of Cycloserine on Promoting the Growth of Beneficial Bacteria

[0021] NA liquid medium: glucose 10g, peptone 5g, beef extract 3g, yeast extract 0.5g, deionized water 1000mL, adjust pH to 7.2-7.4, autoclave at 115°C for 30min.

[0022] NA solid medium is NA liquid medium plus 2-3% (w / w) agar.

[0023] The bacterial strain NJQL-A6 is classified as Ralstonia peeteri, and the preservation number is CGMCC No.6628.

[0024] The NJQL-A6 preserved in glycerol was streaked on the NA solid medium, and cultured at 30°C for 2 days; a single colony on the plate was picked and transferred to the NA liquid medium, and cultured overnight at 30°C and 170rpm. Get the fresh bacterial liquid at 4500rpm, centrifuge at normal temperature for 5min, collect the bacterial cells, wash the bacterial cells with sterile normal saline (0.85% NaCl) for 3 times, remove the culture medium, adjust the concentration of the bacterial liquid with sterile normal saline to 1×10 8 CFU / mL,...

Embodiment 2

[0031] Cycloserine enhances the indoor effect of beneficial bacteria against soil-borne Ralstonia solanacearum

[0032] Material: Ralstonia solanacearum QL-Rs1115 isolated from Qilin Town, Nanjing [1] , its red fluorescent protein marker bacterial strain RS-RFP is the model invading pathogen of the present invention, and this bacterial strain is that the red fluorescent protein gene mCherry is transferred into R. solanacearum QL-Rs1115 through plasmid pYC12, can be in the NA of 30 μ g / mL gentamicin Media plate [2] Grow on it, named RS-RFP. Beneficial bacteria NJQL-A6 screened from Nanjing Qilin Town, can inhibit the growth of Ralstonia solanacearum QL-Rs1115.

[0033] Streak the beneficial bacteria NJQL-A6 preserved in glycerol on the NA solid medium, and place the plate in a 30°C incubator for 2 days until a single colony appears. Pick a single colony on the plate and transfer it to NA liquid medium, and culture overnight at 30°C and 170rpm. Get the fresh bacterial soluti...

Embodiment 3

[0040] Cycloserine enhances potted effect of beneficial bacteria against soil-borne Ralstonia solanacearum

[0041] The beneficial bacterium NJQL-A6 was inserted into the NA liquid medium, and shaken at 30°C and 170rpm for 48h. Get the fresh bacterial solution at 4500rpm, centrifuge at normal temperature for 5min, collect the bacterial cells, wash the bacterial cells for 3 times with sterile normal saline (0.85% NaCl), remove the culture medium, and finally adjust the concentration of the bacterial solution to 1×10 with sterile normal saline. 8 About CFU / mL, obtain the suspension of beneficial bacteria and set aside.

[0042] Ralstonia solanacearum QL-Rs1115 was inserted into NA liquid medium, and cultured with shaking at 30°C and 170rpm for 48h. Get the fresh bacterial solution and centrifuge at 4500rpm at normal temperature for 5min, collect the bacterial cells, wash the bacterial cells for 3 times with sterile normal saline (0.85% NaCl), remove the culture medium, and fina...

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Abstract

The invention discloses an application of cycloserine and beneficial bacteria in preventing and controlling tomato soil-borne ralstonia solanacearum. Cycloserine and beneficial bacteria are applied tosoil by adopting a root irrigation method, wherein the beneficial bacteria are bacterial strains NJQL-A6, the strain is named as Ralstonia pickettii, and is preserved in China General MicrobiologicalCulture Collection Center on September 26, 2012, the address of the preservation unit being Datsun Road, Chaoyang District, Beijing, and the preservation number of the strain being CGMCC No.6628. According to the invention, through a resource regulation and control mode, the soil-borne ralstonia solanacearum inhibition capability of beneficial bacteria can be enhanced; the beneficial bacteria andthe cycloserine are applied at the same time, and the cycloserine is used as a resource substance, so that the growth of the beneficial bacteria can be promoted, and the inhibition capability of thebeneficial bacteria on ralstonia solanacearum can be remarkably improved, the morbidity of tomato ralstonia solanacearum is reduced and tomato soil-borne diseases are prevented and controlled.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to the application of resource-coordinated beneficial bacteria to prevent and control tomato bacterial wilt, and in particular to the application of cycloserine and beneficial bacteria NJQL-A6 to control and control tomato soil-borne bacterial wilt. Background technique [0002] Tomato bacterial wilt is a soil-borne disease caused by Ralstonia solanacearum (referred to as Ralstonia solanacearum). Production reduction restricts the development of agricultural economy. Traditional methods such as application of pesticides or fumigation can kill soil-borne R. solanacearum in a short period of time, effectively slowing down the occurrence of the disease. However, the above methods will also kill the indigenous microorganisms in the soil and reduce the diversity or abundance of soil species. [0003] At present, the application of beneficial bacteria is a relatively green and sustainabl...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/20A01P1/00C12R1/01
CPCA01N63/20C12N1/20C12N1/205C12R2001/01
Inventor 杨天杰韩岗徐阳春韦中沈其荣
Owner NANJING AGRICULTURAL UNIVERSITY
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