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AFLP-STS primer for establishing wheat fingerprint and method for establishing wheat fingerprint by using SSR mark and AFLP-STS mark

An AFLP-STS, wheat technology, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of inability to accurately represent genetic diversity among varieties, inappropriate DNA fingerprinting analysis of a large number of samples, and high DNA quality and quantity requirements. Achieve the effect of low test cost, easy identification and recording, and saving electrophoresis time

Inactive Publication Date: 2010-09-29
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, AFLP is not suitable for DNA fingerprint analysis of a large number of samples due to the many test steps, high requirements for DNA quality and quantity, high test cost, and too many PCR products that are difficult to identify.
In addition, according to our sequencing of some AFLP fragments, some AFLP fragments have the same length, but the DNA sequences are completely different, which proves that these polymorphic fragments do not come from the same DNA site, so only based on the AFLP polymorphic fragments, it is not yet possible Accurately represent genetic diversity among breeds

Method used

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  • AFLP-STS primer for establishing wheat fingerprint and method for establishing wheat fingerprint by using SSR mark and AFLP-STS mark
  • AFLP-STS primer for establishing wheat fingerprint and method for establishing wheat fingerprint by using SSR mark and AFLP-STS mark
  • AFLP-STS primer for establishing wheat fingerprint and method for establishing wheat fingerprint by using SSR mark and AFLP-STS mark

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Establishing Unique DNA Fingerprint Codes for 10 Varieties Planted in Large Areas in my country

[0066] 1. Establish the SSR fingerprint code of wheat varieties

[0067] First, use the above 10 pairs of SSR primers to test the purity of the seeds of each variety, and eliminate hybrids to ensure the correctness of the test samples. Subsequently, the PCR product of each pair of SSR primers of each variety and the control samples of various band types produced by each pair of primers are electrophoresed together, and the SSR band type numbering (table 10, The SSR band pattern number used in the table is a method proposed by the author of this article to quickly and accurately record the SSR band pattern. Method"), according to the fixed sequence of 10 pairs of SSR primers, the band pattern numbers of the tested varieties were gathered to form the SSR fingerprint codes of the varieties.

[0068] Table 10 Band pattern numbers of 10 SSR loci in 10 varieties

[0...

Embodiment 2

[0077] Example 2 Detection of Species Specificity

[0078] The same method as in Example 1 was used to establish DNA fingerprint codes for 140 varieties of the national winter wheat regional test from 2005 to 2006. In the regional test implementation plan for winter wheat varieties) and 105 varieties (variety names are published in the "National Crop Approved Variety List" published by China Agricultural Science and Technology Press) each variety of the approved variety was compared with the remaining 244 varieties, and the following 7 varieties were found: The same or similar DNA fingerprint codes for varieties. In order to calculate the genetic similarity coefficient of each pair of breeds, SSR loci were added for further detection. A total of 98 DNA loci were detected for each pair of breeds. According to the calculation formula of genetic similarity coefficient GS ij =2N ij / (N i +N jj ), calculated the genetic similarity coefficient (N ij is the same allelic variatio...

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Abstract

The present invention relates to a method that the DNA fingerprint of wheat is established by using two molecular markers of SSR and AFLP-STS, and a group of AFLP-STS primers which are used for establishing the DNA fingerprint of wheat. The method comprises SSR fingerprint codes for establishing the breed variety, TS fingerprint codes and steps collecting the fingerprint codes; the STS markers and the SSR markers are distributed in different areas in the wheat genome, and the DNA fingerprint which is established by using two molecular markers can more fully reflect the genetic diversity of the breed variety; the testing steps of the STS markers are as same as the testing steps of the SSR markers, and the testing expense is low; compared with the SSR markers, the belt type of the STS markers which is expanded through the STS primer is simple and easy to be recognized and recorded, according to the product length expanded through each primer, the products of a few of primers are electrophoresed on one gel, the electrophoresis time and the experimental cost are saved, thus not only the ascendance of high polymorphism of an AFLP marker can be kept, but also the default of the tedious recording method of the SSR markers can be overcome.

Description

technical field [0001] The invention relates to a set of AFLP-STS primers for establishing wheat DNA fingerprints and a method for establishing wheat DNA fingerprints using SSR and AFLP-STS molecular markers. Background technique [0002] Scientifically and accurately identifying the genetic differences of wheat varieties and germplasm materials is of great significance for core germplasm selection, new variety identification, and seed purity testing. In the long-term wheat breeding process, the main method to identify the differences between varieties and germplasm resources is to rely on the description of each variety or material phenotype, such as plant height, plant type, ear length, ear shape, leaf color, leaf color, etc. type, grain shape, grain color, and growth period, tillering ability, winter-spring and response to pests and diseases. However, because the above-mentioned agronomic traits are easily affected by the environment, some traits will vary within a certa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 王立新赵昌平张风廷孙辉张立平常利芳黄岚
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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