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Detection method suitable for molecular identification and authentic right identification of crop variety

A technology of identity identification and detection method, which is applied in the field of detection of molecular identity identification and identification of crop varieties, can solve the problems of lack of a comprehensive and systematic detection method, achieve high variety discrimination ability, highlight detection efficiency, and operability strong effect

Active Publication Date: 2017-09-29
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although the molecular identification of crop varieties has more than 20 years of research experience and a rich research foundation, it is widely used in variety management, market monitoring, and judicial identification, but the existing identification standards or technologies are relatively independent, and can only be detected in one or several categories. There is no comprehensive and systematic detection method

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  • Detection method suitable for molecular identification and authentic right identification of crop variety
  • Detection method suitable for molecular identification and authentic right identification of crop variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 The core of maize (diploid plant) variety identification and the acquisition of extended SNP marker site combinations

[0036] 1. Determination of the expansion site combination:

[0037] (1) Maize excellent SNP locus pool: select representative maize inbred line samples at home and abroad (covering 9 types of germplasm groups, as well as tropical, sweet waxy, local varieties, etc.) for deep genome-wide sequencing (10×), from Mining and locating SNP sites in high-quality resequencing data, and determining single-copy and dimorphic SNP sites based on bioinformatics methods; combining SNP sites in published chips or SNP sites in haplotype maps to form relatively A comprehensive maize excellent SNP site pool (the number of sites is greater than 20 million).

[0038] (2) A set of candidate loci suitable for maize variety identification: the above-mentioned excellent SNP locus information was submitted to two American chip production companies, Affymetrix and Illu...

Embodiment 2

[0046] Example 2 The core of wheat (hexaploid) variety identification and the acquisition of extended SNP site combinations

[0047] 1. Determination of the expansion site combination:

[0048] (1) Wheat excellent SNP locus pool: Based on the physical map information of genetic variation loci in the whole wheat genome, comprehensively analyze the distribution, copy number, homology and polymorphism level of SNP loci within and between subgenomes , to screen specific markers that have the ability to identify homologous chromosomes and are highly conserved within cultivars, and establish a pool of excellent marker sites for chromosome-specific SNPs in subgroup A, subgroup B, and subgroup D of hexaploid wheat.

[0049] (2) A set of candidate loci suitable for identification of wheat varieties: the information of the above-mentioned excellent SNP loci was submitted to two American chip production companies, Affymetrix and Illumina, for evaluation. Chip design evaluation and biolo...

Embodiment 3

[0056] The construction of embodiment 3 corn DNA fingerprint database

[0057] (1) Reference varieties: Jing 724 and Jing 92, the two parents of corn hybrid Jingke 968, were selected as reference varieties. The homozygosity of the samples was detected by the combination of extended loci, and the homozygosity reached more than 99.9%.

[0058](2) DNA preparation of maize varieties: each sample was mixed with 30 green leaves of individual plants, ground with liquid nitrogen, and the specific steps of DNA extraction were carried out in accordance with the standards for molecular identification of maize DNA (Wang Fengge et al., 2014). Use ultraviolet spectrophotometer and agarose electrophoresis to detect the quality of the extracted DNA respectively. Agarose electrophoresis shows that the DNA band is single and there is no degradation; the ultraviolet spectrophotometer detects that A260 / 280 is between 1.8-2.0; A260 / 230 is greater than 1.8 . Dilute the DNA to form a working soluti...

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Abstract

The invention discloses a detection method suitable for molecular identification and authentic right identification of a crop variety. Multiple methods and technologies are comprehensively utilized, marker sits undergo omnibearing multi-angle assessment, and a suitable core and expansion site combination is determined to protrude the detection efficiency on the premise of meeting the detection accuracy. The method comprises the following steps: 1, preparing dozens of to hundreds of core site combinations having the characteristics of high stability, high variety distinguishing ability and good compatibility with multiple platforms and being able to highly-efficiently distinguish different varieties; 2, preparing thousands of to tens of thousands of expansion site combinations having the characteristics of uniform distribution and good compatibility with a chip platform and being able to accurately evaluate the heredity similarity among the varieties; and 3, using the two groups of the sites: primarily using the core sites to carry out detection during the identification of the varieties, and adopting the expansion sites to continuously carry out detection if a sample to be detected and a contrast sample cannot be effectively distinguished. The detection method is suitable for the standard DNA fingerprint data construction, molecular identification and authentic right identification of the crop variety.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a detection method suitable for molecular identification and right identification of crop varieties. Background technique [0002] The country is based on agriculture, and agriculture is based on seeds. With the rapid development of breeding and seed industries, the number of crop varieties such as corn, rice, and wheat has increased dramatically, which has brought challenges to both variety identification and seed management. Problems such as counterfeit sets, too much mess, more than one product, and false crowns are prominent, and these problems seriously restrict the development of the modern seed industry. The root of solving these problems lies in the effective identification, monitoring and protection of crop varieties and breeding materials. [0003] Traditional field planting identification is limited by long period, high cost, environmental impact, and dif...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 赵久然王凤格田红丽易红梅王蕊葛建镕
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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