A set of ssr molecular markers for identifying maize varieties and their applications

A molecular marker, corn technology, applied in the field of crop molecular biology, can solve the problems of low accuracy, occupation of land, large workload, etc., and achieve the effects of convenient operation, cost saving and efficiency improvement

Active Publication Date: 2021-04-06
黑龙江省农业科学院农产品质量安全研究所
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These identification methods have application limitations such as long time, heavy workload, low accuracy, and land occupation. At the same time, this method is only suitable for the identification of samples with obvious differences between varieties.

Method used

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  • A set of ssr molecular markers for identifying maize varieties and their applications
  • A set of ssr molecular markers for identifying maize varieties and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Determination of corn SSR molecular markers and primers

[0029] 36 maize varieties were selected for SSR primer screening. Use denaturing polyacrylamide gel electrophoresis to select amplified sites with clear band patterns and high polymorphisms from 987 pairs of SSR primers (https: / / ftp.maizegdb.org / MaizeGDB / FTP / SSRs / ) 50 pairs of primers are available. Using DNA of Heyu 187 and Denghai 18 as templates, 50 pairs of optional primers were used for PCR amplification. According to the results of polyacrylamide gel electrophoresis, the primers with significant differences in the amplified fragments of Heyu 187 and Denghai 18 were screened out.

[0030] Table 1. A set of primer combinations used to identify maize varieties

[0031]

Embodiment 2

[0032] Example 2 Utilize the SSR molecular marker provided by the present invention to distinguish maize varieties

[0033] (1) Rapid extraction of DNA

[0034] DNA was extracted from maize seeds by alkaline boiling method. This method is quick and easy to operate, has no toxic and harmful reagents, is suitable for DNA preparation in the field of plant molecular biology, and is of great significance for greatly shortening the time of seed purity inspection and transgenic detection, improving detection efficiency, and reducing detection costs. The specific operation steps are as follows: take several corn seeds and place them in a 1.5mL centrifuge tube; add 400 μL NaOH (1M) to the centrifuge tube to ensure that the seeds are completely soaked, and bathe in boiling water for 5 min; add 200 μL Tris-HCl ( 1M, pH 8.0), in a boiling water bath for 1 min; add 200 μL TE buffer (pH 8.0), fully dissolve and set aside.

[0035] (2) Quality and quantity of DNA samples

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Abstract

The invention provides a set of SSR molecular markers for identifying maize varieties and its application, a total of two SSR molecular markers, and a set of methods for detecting the SSR molecular markers suitable for a polyacrylamide gel electrophoresis platform. The two SSR molecular markers can be used to quickly distinguish the varieties of corn Heyu 187 and Denghai 18, and realize the combination of SSR markers and high-efficiency and high-sensitivity technology, which saves costs and fully improves the test efficiency in terms of time. Fast, convenient operation, short time, accurate and reliable results and other advantages. The set of primers provided by the invention can be used in the construction of maize fingerprints, identification of varieties, analysis of genetic diversity, etc., and has very broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of crop molecular biology, and in particular relates to a group of SSR molecular markers for identifying maize varieties and applications thereof. Background technique [0002] Corn is an important food crop and feed crop in my country. It is known as a longevity food. It is rich in protein, fat, vitamins, trace elements, etc. It has great potential for developing high-nutrition and high-biological function foods. However, due to its complex heredity and rich variety of variations, the purity and quality of corn seeds are directly related to the yield of crops and the interests of farmers. Improving the purity of corn seeds and regulating the seed market are of great significance to improving grain production capacity and increasing farmers' income. Corn is the third largest food crop in the world. The degree of commercialization of corn seeds is high, and the varieties of corn seeds on the market are mixe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 关海涛张瑞英张晓磊温洪涛李悦
Owner 黑龙江省农业科学院农产品质量安全研究所
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