Streptomyces strains and joint application thereof in prevention and treatment of cucumber fusarium wilt

A technology of streptomyces and streptomyces albicans, applied in the field of microbiology, can solve the problems of ineffective treatment and prevention

Inactive Publication Date: 2015-02-11
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There have been many reports on the biological control of cucumber wilt pathogens by screening antagonistic actinomycetes from soil, but most of them focus on

Method used

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  • Streptomyces strains and joint application thereof in prevention and treatment of cucumber fusarium wilt

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: ( Streptomyces albus 2013 and Streptomyces tenebrarius Isolation and screening of M929)

[0019] Follow these steps:

[0020] (1) Materials and culture medium: The materials used to isolate Streptomyces albicans 2013 and Streptomyces darkens M929 are soil samples collected from the rhizosphere of wetland plants in Xixi, Zhejiang, and the medium used for the isolation of actinomycetes contains dichromic acid Potassium-based PDA medium (the formula is: weigh 200g potatoes, wash, peel and chop them, add 1L of distilled water to boil for half an hour, filter with gauze, add 20g of glucose and 20g of agar, fully dissolve, filter with gauze while hot, divide Pack into conical flask or glass test tube, sterilize under high pressure at 121°C for 20min; add 20μg·mL potassium dichromate when using -1 ; In the following experiments, the medium used for routine culture of actinomycetes was all configured with distilled water);

[0021] (2) Separation and purifi...

Embodiment 2

[0029] Embodiment 2: ( Streptomyces albus 2013 and Streptomyces tenebrarius Preparation method of M929 fermentation metabolite)

[0030] Follow these steps:

[0031] (1) Activation culture of strains: the stored Streptomyces albus 2013 and Streptomyces tenebrariusM929 spore suspension (1×10 8 cfu mL -1 ) into the potato dextrose liquid medium, cultivated for 48 hours at 28°C±1°C for fermentation;

[0032] (2) Fermentation culture: Fermentation medium contains 10g of soybean cake powder, 4g of corn flour, 20g of soluble starch, 5g of glucose, 5g of peptone, and CoCl 2 0.02g, beef extract 5g, CaCO 3 5g, 5g of yeast extract, 50mL of fermentation medium per 300mL triangular bottle; after preparation, sterilize at 121°C for 20min, and use potato glucose liquid medium to culture Streptomyces albus 2013 and Streptomyces tenebrarius M929 was added to the fermentation medium at 5% (v / v) respectively, under the condition of 28°C±1°C, the shaker speed was 180 r / min, a...

Embodiment 3

[0035] Embodiment 3: ( Streptomyces albus 2013 and Streptomyces tenebrarius Inhibitory effect of metabolites of M929 on Fusarium wilt of cucumber)

[0036] (1) Accurately weigh Streptomyces albus 2013 and Streptomyces tenebrarius M929 metabolite 0.1g, add 10mL volume of sterile water, fully dissolve by ultrasonic vibration, respectively prepare the mother solution with a final concentration of 10 g / L;

[0037] (2) Determination of antibacterial activity: Take the above-mentioned mother liquor and dilute it into solutions with a concentration of 1g / L and 0.5g / L respectively, put 1mL of each concentration solution in a sterile petri dish, and rapidly mix ( Streptomyces albus 2013 and Streptomyces tenebrarius The final concentrations of M929 metabolites are respectively 0.1g / L and 0.05g / L), after cooling, put 1 diameter of Cucumber wilt fungus bacterium cake with a diameter of 4mm on each medium plane, and the bacterium cake is connected to the center of the p...

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Abstract

The invention discloses streptomyces strains and joint application thereof in prevention and treatment of cucumber fusarium wilt and relates to the technical field of microbes. Actinomycetes disclosed by the invention are obtained by separation from soil samples collected from rhizospheres of plants in Xixi Wetland, Zhejiang by adopting an actinomycetes separation technology, identified by microbial taxonomy and named as Streptomycesalbus 2013 and Streptomycestenebrarius M929, wherein S. albus 2013 was collected in China Center for Type Culture Collection on June 19, 2013 with the collection number of CCTCC NO: M2013269; and S. tenebrarius M929 was collected in China Center for Type Culture Collection on December 25, 2013 with the collection number of CCTCC NO: M2013710. The two strains can be fermented to produce active substances with relatively strong antagonistic effect against the cucumber fusarium wilt (Fusariumoxysporumf. sp. cucumebrium) and can be used for preventing and treating the cucumber fusarium wilt. When fermentation products of the two strains are mixed according to the ratio in percentage by weight of (3:2)-(1:1) to prepare a mixed preparation, the effect of preventing and treating pathogenic bacteria of the cucumber fusarium wilt is more significant than that when each of the two strains is used as a single preparation respectively.

Description

technical field [0001] The invention relates to the technical field of microorganisms, and relates to the application of metabolites of two strains of biocontrol actinomycetes, Streptomyces albicans 2013 and Streptomyces diablos M929, in the prevention and treatment of cucumber wilt. technical background [0002] Fusarium wilt of cucumber, also known as wilting disease and dead seedling disease, is one of the most serious diseases of cucumber and causes the largest economic loss. In recent years, the incidence of cucumber wilt has increased year by year, which generally leads to a 10% to 20% reduction in cucumber yield, and when the disease is severe, the loss can reach more than 70%, causing huge losses to production. [0003] The pathogen of cucumber wilt is Fusarium oxysporum cucumber-specific Fusarium oxysporum f. sp. Cucumerinum , the pathogen survives the winter in the soil, diseased plant residues, and immature farmyard manure or attached to seeds, and becomes the...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/47C12R1/465
CPCA01N63/10C12N1/205C12R2001/465C12R2001/47
Inventor 马正俞晓平路丹丹罗帅边亚琳许益鹏王正亮申屠旭萍
Owner CHINA JILIANG UNIV
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