Rice Magnaporthe grisea DNA fingerprint belt-type encoding system classifying and analyzing method

A technology of DNA fingerprinting and analysis method, which is applied in the research and analysis of the variability of rice pathogens and the field of research and analysis of genetic traits of rice pathogen variability. It can solve problems such as different calculation formulas and standard naming pedigrees of band patterns, so as to avoid repeated research. , easy to interpret, and easy to compare with each other

Inactive Publication Date: 2005-11-23
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

However, because different researchers did not disclose the corresponding acquisition methods for the Pyricularia oryzae pedigree reported by themselves, and did not name the pedigree according to a unified band type standard, and the software and calculation formulas used are different from each other, so the different places are different from each other. Corresponding findings reported by other researchers are difficult to directly compare

Method used

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Embodiment 1

[0028] The rice blast fungus DNA fingerprint spectrum band type coding program classification and analysis method of the present embodiment is carried out specifically for the rice blast fungus in the japonica rice planting areas in Jiangsu and northern China, and its detailed process is as follows:

[0029] 1. Sampling

[0030] The rep-PCR band type and pedigree research The test materials come from panicle blast samples, which are collected from rice at the panicle stage in the five major rice regions of Jiangsu Province. The samples were isolated from single spores and used for genomic DNA extraction. The sample strains were cultured, sporulated and dried at low temperature and stored on rice straw and filter paper discs. The total number of samples analyzed by rep-PCR band pattern was 296.

[0031] 2. Genomic DNA extraction of Magnaporthe grisea

[0032] 2.1 Extract mycelium

[0033] Use the CTAB / NaCl micro-method to prepare the genomic DNA of Magnaporthe grisea. The m...

Embodiment 2

[0060] The method for classification and analysis of the rice pathogen DNA fingerprint spectrum band type coding program of the present embodiment is carried out specifically for the rice blast fungus in the southern rice region of China, and its detailed process is as follows:

[0061] 1. Sampling—collect 105 test strains from Zhejiang and North China, and isolate and cultivate the strains.

[0062] 1.1 Single spore isolation: Select 2-3 branches of panicle necks with typical symptoms from the standard sample collected, rinse the surface with clean water, soak in a small test tube for 6-8 hours until the standard sample is completely wet, and keep it moist for 24 hours at 26°C. About 1 hour (generally based on the sporulation of lesion spot), put the standard sample on the glass slide of the modified microscope, observe the sporulation situation under the microscope, pick out single spores with a needle fixed on the stage, and place them on the PDA. culture medium (potato, su...

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Abstract

The invention relates to a rice Magnaporthe grisea DNA fingerprint belt-type encoding system classifying and analyzing method, which comprises ill strain acquisition, separating single-spore bacterial strain, extracting genome DNA, expanding rep-PCR, forming fingerprint pattern through electrophoresis, and obtaining encoding pattern based on the fingerprint pattern. The invention realizes fast monitoring for rice bacterium variation by means of molecular genetics, and provides feasible encoding formula for easy reading and comparison of research result at different places.

Description

technical field [0001] The invention relates to a method for researching and analyzing the variability of rice pathogens, in particular to a method for studying and analyzing genetic traits of the variability of rice pathogens, and belongs to the field of agricultural science and technology. Background technique [0002] Rice blast, bacterial blight, and sheath blight are serious hazards in the north and south rice regions of my country, and are the three major diseases of rice in my country. Among them, rice blast caused by Magnapor the grisea is the most widespread and harmful worldwide rice disease. Breeding and utilizing disease-resistant varieties is currently the most effective and practical control method. However, due to the serious differentiation of pathogenicity of Magnaporthe oryzae, various physiological races, large pathogenicity changes, and easy mutation, a highly resistant variety often becomes a highly susceptible variety within 3 to 5 years. The selectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00
Inventor 周益军程榜范永坚白娟
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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