Method for direct extraction of pathogen DNA from pathogenic tissue of Graminaceous crop

A technology of gramineous crops and pathogenic bacteria, which is applied in the field of microbial molecular detection, can solve the problems of unfavorable sample analysis and detection, long experiment cycle, high cost, etc., and achieve the effect of reducing tediousness, rapid detection and monitoring, and shortened time

Inactive Publication Date: 2015-04-08
INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The CTAB method requires the grinding of liquid nitrogen in the early stage, or the use of enzymes to break the cell wall, and then uses toxic reagents such as chloroform to extract to remove proteins, DNA precipitation and elution and other steps. Although high-quality DNA can be obtained, but The whole process is time-consuming and inefficient
Moreover, the isolation and cultivation of pathogenic microorganisms need to be completed in the early stage of DNA isolation, and the experiment cycle is long.
Although the method of kit extraction is much simpler than the traditional method, it has the problem of high cost, which is not conducive to the analysis and detection of a large number of samples

Method used

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  • Method for direct extraction of pathogen DNA from pathogenic tissue of Graminaceous crop
  • Method for direct extraction of pathogen DNA from pathogenic tissue of Graminaceous crop
  • Method for direct extraction of pathogen DNA from pathogenic tissue of Graminaceous crop

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Embodiment Construction

[0024] It is to illustrate how the present invention is carried out by means of examples. These examples are only to illustrate how the present invention is realized, or the best implementation mode. These examples can not limit the present invention in any way. The scope of the present invention is the claims limited.

[0025] Implementation Example 1: Extraction and verification of DNA from rice leaves and Magnaporthe grisea.

[0026] The specific implementation method:

[0027] 1) Reagents: Prepare the following reagents in advance and use them after sterilization: (1), 100mM TrisHCl (pH8.7), (2), 10mM EDTA (pH 8.0), (3, )1M KCl, (4), 10% Tween 20.

[0028]2) Sample preparation: randomly select 15 typical diseased leaves and panicles that were collected from the field and stored in the laboratory at room temperature, cut diseased leaves with a size of about 2 mm × 2 mm from diseased rice leaf lesions, and cut diseased leaves from diseased rice leaves. Take 1-2mm long pan...

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Abstract

The invention provides a method for direct extraction of pathogenic fungus and bacteria DNA from leaf scabs of gramineous crops such as pathogenetic rice and corn. The method of the invention can directly extract pathogen DNA from leaf and neck of rice and corn leaf scabs infected by rice blast, the extraction method is simple, fast and low-cost, and the extracted DNA is applicable to PCR based analysis on pathogenic organism molecular detection and diagnosis, pathogen nontoxic gene detection and colony genetic diversity.

Description

technical field [0001] The invention belongs to the field of microbial molecular detection, in particular to a method for directly extracting DNA of pathogenic bacteria from susceptible tissues of gramineous crops. Background technique [0002] The extraction of DNA from crop pathogenic microorganisms is the basis for the detection and analysis of related pathogenic molecules. The methods commonly used to extract the DNA of traditional pathogenic fungi are CTAB extraction and cell wall lysis. The CTAB method requires the grinding of liquid nitrogen in the early stage, or the use of enzymes to break the cell wall, and then uses toxic reagents such as chloroform to extract to remove proteins, DNA precipitation and elution and other steps. Although high-quality DNA can be obtained, but The whole process is time-consuming and inefficient. Moreover, the isolation and cultivation of pathogenic microorganisms need to be completed in the early stage of DNA isolation, and the exper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68C12Q1/04
Inventor 董丽英杨勤忠刘树芳徐鹏李静
Owner INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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