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2RL chromosome-specific codominant KASP molecular mark for rye and its application

A molecular marker and chromosome technology, applied in the fields of biotechnology and genetic breeding, to achieve the effects of good genetic stability, high resolution and high conservation

Active Publication Date: 2019-01-18
中国科学院遗传与发育生物学研究所农业资源研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no co-dominant KASP markers have been reported for the detection of outer margin rye chromosomes in wheat background

Method used

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  • 2RL chromosome-specific codominant KASP molecular mark for rye and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Obtainment of rye universal 2RL chromosome-specific co-dominant KASP molecular marker SWK2RL1

[0030] 1. Genomic DNA extraction and purification

[0031] Genomic DNA of five types of rye, including imperial rye, German white rye, Jingzhou rye, hundred-grain rye and Guyuan rye, were extracted by CTAB method. The specific operation steps were as follows:

[0032] (1) Take the fresh leaves of two leaves and one heart stage in a 2mL centrifuge tube, freeze them in liquid nitrogen and grind them into powder;

[0033] (2) Add 600 μL of CTAB extract preheated at 65°C to the centrifuge tube, bathe in water at 65°C for 40 minutes, and shake gently every 10 minutes to mix;

[0034] (3) Take out the centrifuge tube and cool it down for a few minutes, add chloroform-isoamyl alcohol (24:1) equal to the CTAB extract to the centrifuge tube, place on a shaker for 15 minutes to shake well, centrifuge at 12000rpm for 10 minutes, take Transfer the supernatant to a new 1.5mL c...

Embodiment 2

[0057] Example 2 Application of rye universal 2RL chromosome-specific co-dominant KASP molecular marker SWK2RL1

[0058] Materials to be tested include:

[0059] 8 wheat varieties: Zhongguochun, Xiaoyan 6, Holdfast, Kenong 199, Gaocheng 8901, Jishi 02-1, Shixin 633, Shixin 733;

[0060] 7 rye varieties: Imperial rye, German white rye, Jingzhou rye, hundred-grain rye, Guyuan rye, Weining rye, KingII rye;

[0061] 30 wheat-rye materials identified by genome in situ hybridization GISH / FISH and confirmed the presence of outer border chromosomes: Holdfast-KingII wheat-rye complete set of disomic addition lines and diplosome addition lines, a total of 21, China Spring-Imperial There are 7 rye 1R-7R addition lines in total, Xiaoyan 6-German white rye 2RL double-end addition line WR56, Xiaoyan 6-German white rye 2R(2D) disomic substitution line WR91.

[0062] Using the DNA extracted from the above materials as a template, use the primers for the KASP molecular marker SWK2RL1 develop...

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Abstract

A 2RL chromosome specific codominant KASP molecular mark for rye and its application are disclosed. That KASP molecular mark is SWK2RL1, and the primer of the KASP molecular mark SWK2RL1 comprises anallelic upstream primer of rye, an allelic upstream primer of wheat and a common downstream primer. The nucleotide sequence of the rye allelic upstream primer is shown in SEQ ID NO: 1; the nucleotidesequence of the wheat allelic upstream primer is shown in SEQ ID NO: 2; the nucleotide sequence of the downstream primer is shown in SEQ ID NO: 3. The invention is based on the difference SNP site between rye and wheat, The codominant KASP marker SWK2RL1 of rye universal 2RL chromosome was developed for the first time. This marker has good genetic stability and high resolution, and can be used inwheat chromosome engineering breeding and molecular marker assisted selection breeding. It provides a tool for high throughput detection and tracing of rye chromosome 2RL from different sources in wheat background and accelerates the utilization and transfer of excellent genes on rye chromosome 2RL.

Description

technical field [0001] The invention relates to the fields of biotechnology and genetic breeding, in particular to a rye universal 2RL chromosome-specific co-dominant KASP molecular marker and its application. Background technique [0002] Wheat is one of the most important food crops in the world. The high and stable yield of wheat plays a vital role in the development of national economy and national food security. However, with the increase in the number of crosses between varieties in the long-term traditional breeding process, the high genetic consistency among the main varieties limits the further improvement of wheat yield, quality and resistance, making it difficult to achieve greater breakthroughs in wheat genetic improvement. Close relatives of wheat contain rich genetic variation and carry a large number of excellent genes such as high yield, high quality, and resistance to biotic and abiotic stress. Therefore, it has become an urgent need for wheat genetic impro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 安调过韩帼豪靳玉丽王婧刘士毓麻菲菲许云峰
Owner 中国科学院遗传与发育生物学研究所农业资源研究中心
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