A method for identifying the purity of Jingke 968 corn hybrid based on SNP markers

A corn and molecular marker technology, applied in the field of crop molecular biology, can solve the problems of heavy workload, difficult high-throughput detection methods, and long identification period

Active Publication Date: 2022-07-01
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reported methods have problems such as long identification cycle, heavy workload, easy to be affected by environment or developmental stage, and difficult to form high-throughput detection methods

Method used

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  • A method for identifying the purity of Jingke 968 corn hybrid based on SNP markers
  • A method for identifying the purity of Jingke 968 corn hybrid based on SNP markers
  • A method for identifying the purity of Jingke 968 corn hybrid based on SNP markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Determination of SNP molecular markers and primers for identifying the purity of Jingke 968 corn hybrids

[0038] (1) Basic site: Using the 384 SNP sites published in the "Maize DNA Fingerprint Library and Variety Molecular Identification SNP Core Site Combination-maizeSNP384" (published in Chinese Patent 201410756086.0) as the starting site set, and established SNP-DNA fingerprint data of 335 maize hybrids under national review.

[0039] (2) SNP site test and biological characteristics evaluation: 192 corn hybrids were selected, and DNA was extracted by rapid method, CTAB, kit, and various methods. Based on the KASP technology system, the DNA extracted by different methods was used to verify the site. The typing effect, repeatability, and stability of the points are used to obtain a set of candidate sites.

[0040] (3) Evaluation of SNP loci polymorphism: using data of 200 different types of maize inbred lines and 335 maize hybrids, the polymorphism paramete...

Embodiment 2

[0049] Embodiment 2 utilizes the SNP molecular marker provided by the present invention to identify the method for the purity of Jingke 968 corn hybrid

[0050] Using the primer combinations designed in Example 1 for the 4 SNP sites, the scheme for identifying the seed purity of Jingke 968 is as follows: Synthesize primers according to the KASP technical requirements, and the primers are common primers without fluorescent groups; PCR amplification system MasterMix; configure the reaction system to add DNA, primers and MasterMix; run the amplification reaction program; scan the fluorescence signal in situ; perform data analysis to obtain genotype data; The genotype data is used to determine whether the individual plant is a normal plant, an inbred seedling, or an allotype plant, and finally its purity value is comprehensively determined. The specific operations are as follows:

[0051] (1) Preparation of the sample to be tested:

[0052] At least 150 seeds are randomly select...

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Abstract

The invention provides a method for identifying the purity of Jingke 968 corn hybrid based on SNP markers. Based on 384 SNP sites, the invention adopts a high-throughput KASP technology platform, and uses more than 500 samples to conduct test effects and biological characteristics. As well as comprehensive evaluation of polymorphism parameters, 4 SNP markers with high quality, high stability, high polymorphism and high heterozygosity are finally determined, and 3 primers for each SNP marker are designed to amplify a total of 12 primers. The nucleotide sequences are respectively as SEQ ID NO.1-12. The four pairs of primer combinations can be used to identify the seed purity of Jingke 968, and can identify self-crossed seedlings, backcrossed seedlings and heterotypic strains. The invention expands the type and method of identification markers for the purity of corn hybrids, and provides the promotion and planting of Jingke 968. strong technical support.

Description

technical field [0001] The invention belongs to the technical field of crop molecular biology, in particular to a method for identifying the purity of Jingke 968 corn hybrid based on SNP markers. Background technique [0002] Seeds are the basic means of production in agriculture, and the yield-increasing potential of fine varieties is easily affected by the purity of seeds. Purity identification is one of the indicators to determine whether the quality of commercial seeds is qualified, and it is also an important means to prevent fake and inferior seeds from entering the market. The establishment of a fast, efficient, low-cost, and high-throughput identification method for seed purity is very beneficial to improving seed quality monitoring, promoting standardized production and operation, and safeguarding the vital interests of seed industry enterprises and farmers. [0003] The traditional field identification method needs to observe the specific characteristics of variet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/172
Inventor 田红丽王蕊易红梅王元东
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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