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A method for identifying the purity of Jingnongke 728 corn hybrid based on SNP markers

A technology of hybrid purity and corn, applied in the field of crop molecular biology, can solve the problems of difficult high-throughput detection method, long identification period, large workload, etc.

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 Jingnongke 728 corn hybrid based on SNP markers
  • A method for identifying the purity of Jingnongke 728 corn hybrid based on SNP markers
  • A method for identifying the purity of Jingnongke 728 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 Jingnongke 728 corn hybrids

[0038] (1) Basic site: Using the 384 SNP sites published in "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) SNP locus polymorphism evaluation: using 200 different types of maize inbred lines and 335 maize hybrids data, the polymorphism parameters PI...

Embodiment 2

[0049] Embodiment 2 utilizes the SNP molecular marker provided by the invention to identify the method for the purity of Jingnongke 728 corn hybrid

[0050] Using the primer combinations designed in Example 1 for the 4 SNP sites, the scheme for identifying the seed purity of Jingnongke 728 is as follows: Synthesize primers according to KASP technical requirements, and the primers are common primers that do not contain fluorescent groups; purchase KASP technical support 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; Based on the genotype data on it, it is determined 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 test...

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Abstract

The invention provides a method for identifying the purity of Jingnongke 728 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 carry out test effects, biological The comprehensive evaluation of characteristics and polymorphism parameters finally determined 4 SNP markers with high quality, high stability, high polymorphism and high heterozygosity, and designed 3 primers to amplify each SNP marker, a total of 12 primers. The nucleotide sequences are shown in SEQ ID NO.1-12, respectively. The four pairs of primer combinations can be used to identify the seed purity of Jingnongke 728, 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, which is the popularization of Jingnongke 728. Planting provides a strong technical guarantee.

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 Jingnongke 728 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 va...

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 田红丽王蕊杨扬易红梅王元东
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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