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Method for rapid separation of pathogen DNA from rice blast single scab

A rice blast fungus and rice blast technology, applied in the field of microbial molecular detection, can solve cumbersome problems such as grinding with liquid nitrogen, or cell wall-breaking lysis and repeated extraction, long experimental period, and high experimental cost. The effect of reducing cumbersomeness, rapid detection and monitoring, and simplification of procedures

Inactive Publication Date: 2015-04-08
INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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Problems solved by technology

However, using these methods to extract DNA requires several days of preparation in the early stage, such as the isolation, preservation, and cultivation of pathogenic bacteria, resulting in a long experimental period for early preparation; and the process of using these methods to extract DNA is also relatively cumbersome, such as Liquid nitrogen grinding, or cell wall lysis and repeated extraction are required, and some methods, such as using kits to extract DNA, have disadvantages such as high experimental costs, which are difficult to meet the large-scale and rapid development of field pathogenic bacteria groups based on PCR technology. Purpose of detection analysis

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  • Method for rapid separation of pathogen DNA from rice blast single scab

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Embodiment Construction

[0017] 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.

[0018] Implementation Example 1: Extraction and verification of DNA from rice leaves and Magnaporthe grisea.

[0019] The specific implementation method:

[0020] 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.

[0021] 2) Sample preparation: Randomly select 15 typical diseased leaves and panicles that were collected from the field and stored in the laboratory after being dried at room temperature, and cut diseased leaves with a size of about 2 mm × 2 mm from a single lesion of the diseased rice leaf. Take 1-2mm long diseased panicle...

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Abstract

The invention provides a method for direct extraction of rice blast pathogen DNA from rice leaf infected by rice blast or neck blast single scab. The method of the invention can directly extract rice blast pathogen DNA from rice leaf or neck infected by rice blast, the extraction method is simple, fast and low-cost, and the extracted DNA is applicable to PCR based analysis on rice blast 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 rapidly isolating pathogenic bacteria DNA from a single scab of rice blast. Background technique [0002] Rice blast, caused by the fungus Magnaporthe oryzae (Anamorph: Pyricularia oryzae) (Couch and Hohn, 2002), is one of the major limiting factors of rice production, seriously threatening the world's food production (Ou, 1985; Zeigler et al .,1994). The interaction between rice and Magnaporthe grisea conforms to the classical "gene-to-gene" theory (Flor, 1971; Jia et al., 2000). Studying the pathogenic composition of the blast fungus population will help guide the promotion and distribution of new rice varieties resistant to disease. With the deepening of research on the pathogenicity of Magnaporthe grisea, the avirulent genes Avr1Pi-CO39, ACE1, AvrPiz-t, AvrPita, AvrPik / Pikm / kp, AvrPia and AvrPii, etc. Avirulent genes have continued to be successful...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68C12Q1/04C12R1/645
Inventor 杨勤忠董丽英刘树芳徐鹏李静
Owner INST OF AGRI ENVIRONMENT & RESOURCES YUNNAN ACAD OF AGRI SCI
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