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Method for identifying purity of Jingke 968 corn hybrids based on SNP markers

A technology for hybrid purity and corn, applied in the field of molecular biology of crops, can solve the problems of heavy workload, easy to be affected by environment or developmental stage, and difficult to high-throughput detection methods.

Active Publication Date: 2020-03-10
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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  • Method for identifying purity of Jingke 968 corn hybrids based on SNP markers
  • Method for identifying purity of Jingke 968 corn hybrids based on SNP markers
  • Method for identifying purity of Jingke 968 corn hybrids based on SNP markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Determination of SNP molecular markers and primers used to identify the purity of Jingke 968 corn hybrids

[0038] (1) Basic sites: The 384 SNP sites published by "Maize DNA Fingerprint Library and Variety Molecular Identification SNP Core Site Combination-maizeSNP384" (disclosed in Chinese Patent No. 201410756086.0) were used as the starting site set and established SNP-DNA fingerprint data of 335 nationally approved corn hybrids.

[0039] (2) SNP site test and biological feature evaluation: 192 corn hybrids were selected, and DNA was extracted by rapid extraction, CTAB, kits, and multiple methods. Based on the KASP technology system, the DNA verification sites extracted by different methods were used. The typing effect, repeatability, and stability of the points can be used to obtain a set of candidate sites.

[0040] (3) Polymorphism evaluation of SNP locus: using 200 different types of maize inbred lines and 335 maize hybrids data to evaluate the polymorphism pa...

Embodiment 2

[0049] Example 2 Using the SNP molecular marker provided by the present invention to identify the purity of Jingke 968 corn hybrid

[0050] Using the primer combination designed for 4 SNP sites in Example 1, the scheme for seed purity identification of Jingke 968 is as follows: Synthesize primers according to the KASP technical requirements, and the primers are common primers without fluorescent groups; purchase KASP technology matching 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; according to each individual plant of Jingke 968 on each pair of primers Genotype data, determine whether the individual plant is a normal plant, a selfed seedling, or a heterogeneous plant, and finally determine its purity value comprehensively. The specific operations are as follows:

[0051] (1) Preparation of samples to be...

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Abstract

The invention provides a method for identifying the purity of Jingke 968 corn hybrids based on SNP markers. The method is based on 384 SNP loci, a high-flux KASP technology platform is adopted, more than 500 samples are subjected to comprehensive evaluation of test effects, biological characteristics and polymorphic parameters, finally, four SNP markers with high quality, high stability, high polymorphism and high heterozygosis rate are determined, three primers of each SNP marker are designed and amplified, and the 12 primers have the nucleotide sequences shown as SEQ ID NO.1-12 respectively.By utilizing the four pairs of primer combination, the seed purity of Jingke 968 can be identified, selfing seedlings, backcross seedlings and special-shaped plants can be identified, the types and methods of corn hybrid purity identification markers are expanded, and a powerful technical guarantee is provided for popularization and planting of Jingke 968.

Description

Technical field [0001] The invention belongs to the technical field of crop molecular biology, and specifically relates to a method for identifying the purity of Jingke 968 corn hybrids based on SNP markers. Background technique [0002] Seeds are the basic means of production in agriculture, and the yield-increasing potential of good varieties is easily affected by seed purity. Purity identification is one of the indicators to determine whether the quality of commercial seeds is qualified. It is also an important means to prevent fake and inferior seeds from entering the market, and it is also an important way for seed companies to control product quality. The establishment of a fast, efficient, low-cost, and high-throughput seed purity identification method is very helpful to improve seed quality monitoring, promote standardized production and management, and protect the vital interests of seed industry enterprises and farmers. [0003] Traditional field identification methods n...

Claims

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Application Information

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Patent Type & Authority Applications(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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