ISSR fingerprint construction method for identifying reality of rice variety

A technology of fingerprint map and construction method, which is applied in the field of authenticity identification of germplasm resource varieties, can solve the problems of simple and rapid molecular marker identification that are not suitable for the authenticity of crop varieties, and achieve the effect of stable results.

Inactive Publication Date: 2011-01-26
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

[0004] Most of the conventional methods for constructing DNA fingerprints select a representative part of the bands amplified by multiple primers, and use a matrix of "0" and "1" to construct the fingerprints of varieties or germplasm resources, which is not suitable for the production of crop varieties. Simple and rapid molecular marker identification for authenticity

Method used

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  • ISSR fingerprint construction method for identifying reality of rice variety
  • ISSR fingerprint construction method for identifying reality of rice variety
  • ISSR fingerprint construction method for identifying reality of rice variety

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0057] In this example, the seeds of 24 rice varieties mainly planted in Zhejiang Province were used as test materials, the total DNA of the seedling genome was extracted as a template, ISSR-PCR amplification with specified primers was carried out, and 1.5% agarose gel electrophoresis was performed on the ISSR-PCR products Detect and take pictures, and establish the ISSR molecular marker fingerprints of each test sample according to the electrophoresis bands, and use the constructed ISSR fingerprints to identify the authenticity of rice varieties.

[0058] The specific operation steps are as follows:

[0059] (1) DNA extraction

[0060] The seeds of 24 rice varieties mainly planted in Zhejiang Province were used as materials, and the young leaves were taken at the seedling stage to extract DNA by the improved CTAB method, and a part of the DNA solution was taken to measure its concentration, diluted to 20ng / μL, and stored at -20°C for later use;

[0061] (2) ISSR-PCR amplific...

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Abstract

The invention discloses an ISSR fingerprint construction method for identifying the reality of a rice variety and provides a quick and accurate rice variety ISSR fingerprint construction method and use thereof. The method comprises the following steps: extracting DNA of young rice seedlings by an improved CTAB method; performing ISSR-PCR amplification by using the DNA of the young seedlings as a template; performing the 1.5-percent agarose elelectrophoretic detection of the product of the PCR, and taking an image in ultraviolet light; and constructing the DNA fingerprint of the rice variety for identifying the reality of the rice variety. In the invention, the DNA fingerprint contains basic information including rice variety code, amplified band types, site code of each band and '0' and '1' arrays. The method has the characteristics of simple operation, visualization, practicality and the like and promotes the wide use of the ISSR molecular mark technology in the accurate, quick, simple and convenient identification of the reality of the rice variety.

Description

technical field [0001] The invention relates to a germplasm resource variety authenticity identification technology, and is an ISSR fingerprint spectrum construction method suitable for rice variety authenticity identification. Background technique [0002] Rice is an important food crop. my country is the largest producer and consumer of hybrid rice seeds in the world, and has a huge market for hybrid rice seeds. The authenticity and purity of rice seeds are important indicators to measure the quality of seeds, which directly affect the yield, quality and interests of farmers. How to quickly, accurately, simply and economically identify the authenticity of hybrid rice seeds in production is a prominent problem. The traditional morphological identification method relies on the phenotypic differences of varieties, and the performance of morphological traits is greatly affected by the environment, and the morphological identification method has a large workload, long identifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 胡晋朱岩芳关亚静祝水金
Owner ZHEJIANG UNIV
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