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Method for simply, conveniently and rapidly separating peronophthora litchii

A technology of litchi downy mildew and litchi skin, applied in the direction of using spores, methods based on microorganisms, and separating microorganisms, can solve the problems of cumbersome operation and difficult success, and achieve simple operation steps, improved efficiency, and improved coverage and biologically active effects

Active Publication Date: 2021-11-12
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, conventional methods for isolating fungi cannot be used to separate Phytophthora litchie well, and the operation is cumbersome and difficult to succeed.

Method used

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  • Method for simply, conveniently and rapidly separating peronophthora litchii
  • Method for simply, conveniently and rapidly separating peronophthora litchii

Examples

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Effect test

Embodiment 1

[0024] A method for isolating downy mildew of litchi, comprising the steps of:

[0025] (1) Take litchi diseased fruit, cut off the damaged part as the diseased tissue, rinse with water, add 70% ethanol solution with volume concentration to soak for 0.5min, rinse with sterile water for 3 times, and dry the surface moisture;

[0026] (2) By weight, get V8 vegetable juice 80g, add water 850g, after sterilization and cooling, get V8 vegetable medium, add ampicillin 0.1g, prochloraz 0.1g and hymexazol 0.1g, add growth promoting 40g of β-sitosterol mother liquor, 40% of lychee bark research solution and the balance are mixed with water in terms of mass percentage, to obtain selective culture medium; 4g of β-sitosterol mother liquor, 16g of lychee peel extract and 20g of water.

[0027] The β-sitosterol mother liquor is obtained by mixing and dissolving 10g of β-sitosterol and 1g of dichloromethane at a mass ratio of 10:1; the litchi skin research solution is obtained by mass ratio...

Embodiment 2

[0031] A method for isolating downy mildew of litchi, comprising the steps of:

[0032] (1) Take the diseased litchi fruit, cut off the damaged part as the diseased tissue, rinse with water, add 70% ethanol solution to soak for 1 min, rinse with sterile water for 3 times, and dry the surface moisture;

[0033] (2) By weight, get V8 vegetable juice 120g, add water 950g, after sterilization and cooling, get V8 vegetable medium, add ampicillin 0.2g, prochloraz 0.2g and hymexazol 0.2g, add growth promoting 80g of β-sitosterol mother liquor, 50% of lychee bark grinding solution and the balance are mixed with water in terms of mass percentage, the growth-promoting agent is mixed to obtain selective medium; 12g of β-sitosterol mother liquor, 40g of lychee peel extract and 28g of water.

[0034] The β-sitosterol mother liquor is obtained by mixing and dissolving 15g of β-sitosterol and 2g of dichloromethane with a mass ratio of 15:2; the lychee skin research solution is based on the ...

Embodiment 3

[0038] A method for isolating downy mildew of litchi, comprising the steps of:

[0039] (1) Take litchi diseased fruit, cut off the damaged part as the diseased tissue, rinse with water, add 70% volume concentration ethanol solution to soak for 0.8min, rinse with sterile water for 3 times, and dry the surface moisture;

[0040] (2) By weight, get V8 vegetable juice 100g, add water 900g, after sterilization and cooling, get V8 vegetable medium, add ampicillin 0.15g, prochloraz 0.15g and hymexazol 0.15g, add growth promoting 70g of β-sitosterol mother liquor, 45% of lychee bark research solution and the balance are mixed with water in terms of mass percentage, to obtain selective culture medium; 9.8g of β-sitosterol mother liquor, 31.5g of lychee skin research solution and 28.7g of water.

[0041] The β-sitosterol mother liquor is obtained by mixing and dissolving 11g of β-sitosterol and 1g of dichloromethane according to the mass ratio of 11:1; the litchi skin research solutio...

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Abstract

The invention provides a method for simply, conveniently and rapidly separating peronophythora litchii. The method comprises the following steps: (1) selecting and disinfecting disease tissues of litchi; (2) preparing a V8 vegetable culture medium, adding benzyl ampicillin, prochloraz, hymexazol and a growth promoting agent, and mixing evenly to obtain a selective culture medium, wherein the growth promoting agent is formed by mixing beta-sitosterol mother liquor, litchi rind ground-out liquid and water; (3) culturing the disease tissues in the selective culture medium, selecting hyphae, conducting microscopic examination, and inoculating the hyphae to a new selective culture medium for culturing; and (4) cutting hypha blocks from a microorganism colony, adding the litchi rind ground-out liquid for soaking, and culturing in a new V8 vegetable culture medium to obtain peronophythora litchii. The peronophythora litchii obtained through separation has good activity and pathogenicity. The method is easy to operate, has a high separation rate, is easy for separation, and can realize efficient separation of the peronophythora litchii.

Description

technical field [0001] The invention relates to the technical field of plant pathogenic bacteria isolation, in particular to a simple and rapid method for isolating Pyrethora peronosa of litchi. Background technique [0002] Litchi frost blight (formerly known as litchi downy mildew or litchi downy mildew) caused by Peronophythora litchii Chen ex Ko et al seriously threatens the healthy development of litchi industry in my country, and is the most harmful to litchi production and postharvest. The most serious and widespread disease. The disease can infect young leaves, twigs, spikes and fruits, causing a large number of spikes and fruits to rot. It can cause 10%-30% yield loss in general years, and 80% yield loss in popular years, especially serious in Guangdong, Guangxi, Fujian, Hainan and other places. In order to conduct related research on litchi downy mildew, it is necessary to obtain pure downy Phytophthora litchie through isolation. There are often other miscellaneo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/38C12N1/14C12N3/00C12R1/645
CPCC12N1/02C12N1/38C12N1/14C12N3/00Y02A50/30
Inventor 陈庆河杨成东贾斯蒂
Owner HAINAN UNIVERSITY
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