Molecular marker and specific primer for identifying wheat grain germination traits and application thereof

A specific primer pair and wheat technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of inability to co-segregate target genes, long distances of target genes, molecular marker-assisted selection bias, etc. question

Active Publication Date: 2016-08-17
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used molecular marker types include AFLP, SSR, etc., but these marker types are often far away from the target

Method used

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  • Molecular marker and specific primer for identifying wheat grain germination traits and application thereof
  • Molecular marker and specific primer for identifying wheat grain germination traits and application thereof
  • Molecular marker and specific primer for identifying wheat grain germination traits and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Discovery of alleles and development of primer pairs

[0058] On the basis of sequence analysis of a large number of wheat varieties, it was found that the wheat TaSdr-A1 gene, two allelic variant forms of the TaSdr-A1 gene in wheat, one as shown in sequence 1 of the sequence table (named allele gene TaSdr-A1a), and the other is shown in sequence 2 of the sequence listing (named allele TaSdr-A1b). The comparison spectrum of the two allelic variant forms of the TaSdr-A1 gene is shown in figure 1 , the start codon and stop codon are shown in boxes, and the underlined part is the coding region.

[0059] Based on the two allelic variations of the TaSdr-A1 gene, specific primer pairs were designed as follows (the target sequence is 1294bp):

[0060] Primer F (sequence 3 of the sequence listing): 5'-CGTCGGCAGACATCGACTCC-3';

[0061] Primer R (SEQ ID NO: 4 of the Sequence Listing): 5'-GAAGCTCACTAGCTCAGAACACGC-3'.

Embodiment 2

[0062] Embodiment 2, the application of allele and primer pair

[0063] The experimental materials were 144 Chinese winter wheat main varieties (National Wheat Improvement Center, Institute of Crop Science, Chinese Academy of Agricultural Sciences), see Table 1 and Table 2 for details.

[0064] 1. Detection of wheat genotype

[0065] Each experimental material was carried out as follows:

[0066] 1. Genomic DNA extraction.

[0067] 2. Using the genomic DNA extracted in step 1 as a template, perform PCR amplification with the specific primer pair designed in Example 1 to obtain a PCR amplification product.

[0068] PCR amplification reaction system (50 μl): template (DNA content about 100ng), ExTaq enzyme (Takara, code: DRR100B) 0.3 μl, GC buffer Ⅰ (Takara, code: DRR20GC I) 25 μl, dNTP (Takara, code: DRR100B) D4030RA, 2.5mM each) 4 μl, primer F (20 μM) 1 μl, primer R (20 μM) 1 μl, and 50 μl of the reaction system was supplemented with sterile ultrapure water.

[0069] The r...

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Abstract

The invention discloses a molecular marker and a specific primer for identifying wheat grain germination traits and application thereof. The invention provides a specific primer pair. The specific primer pair comprises two primers; amplification products comprise specific DNA molecules; the specific DNA molecules are target sequences of a primer F and a primer R in a wheat genome; the primer F is a single-chain DNA molecule shown as a sequence 3 of a sequence table; the primer R is a single-chain DNA molecule shown as a sequence 4 of the sequence table. The invention also provides a molecular marker related to a wheat germination index; the molecular marker is a DNA fragment prepared by taking the genome DNA of the to-be-detected wheat as a template and amplifying by using the specific primers. The molecular marker and the specific primer have important theoretical significance and economic value for assistant selection of wheat variety with preharvest sprouting resistance through the molecular marker.

Description

technical field [0001] The invention relates to molecular markers and specific primer pairs for identifying germination traits of wheat grains and their application. Background technique [0002] Ear sprouting (PHS) refers to the germination of wheat on the ear when it encounters rain or humidity before harvest, and it is relatively common in crops such as wheat, corn, and rice. Ear germination will seriously affect the yield and processing quality of wheat, and the loss is closely related to the severity of ear germination. Because ear germination will reduce the thousand-grain weight of wheat, the yield of wheat fields with ear germination generally decreases by 6-10%, and severely germinated wheat can only be used for livestock feed and cannot be used for flour processing. The activities of amylase, protease and lipase in germinated grains are greatly increased, resulting in the degradation of starch, protein and fat. The strength of flour processed from germinated whea...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 何中虎张颖君夏先春
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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