Primer pair for assaying germination traits of wheat seed grains and related molecular marker

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 wheat ear germination, loss of edible value, inability to co-segregate target genes, etc., to achieve The effect of accelerating the pace of breeding

Inactive Publication Date: 2014-08-06
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

In 2008, serious wheat ear germination occurred in Hebei, Beijing and other places, and the affected area reached hundreds of thousands of mu, and the severe ones had lost their edible value.
Commonly used molecular marker types include RFLP, AFLP, SSR, etc., but these marker types cannot co-segregate with the target gene

Method used

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  • Primer pair for assaying germination traits of wheat seed grains and related molecular marker
  • Primer pair for assaying germination traits of wheat seed grains and related molecular marker
  • Primer pair for assaying germination traits of wheat seed grains and related molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Discovery of alleles and development of primer pairs

[0050] On the basis of sequence analysis of a large number of wheat varieties, it was found that the wheat TaSdr-B1 gene, two allelic variant forms of the TaSdr-B1 gene in wheat, one as shown in sequence 1 of the sequence table (named allelic gene TaSdr-B1a), and the other is shown in sequence 2 of the sequence listing (named TaSdr-B1b). The comparison spectrum of the two allelic variant forms of the TaSdr-B1 gene is shown in figure 1 , the start codon and stop codon are shown in boxes, the underlined part is the coding region, and the shaded part is the sequence polymorphic site.

[0051] Design specific primer pairs based on the two allelic variants of the TaSdr-B1 gene as follows:

[0052] Upstream primer (sequence 3 of the sequence listing): 5'-CGCCTACGTGTCGGCCC-3';

[0053] Downstream primer (sequence 4 of the sequence listing): 5'-TCCGTGACGACCGCCGGG-3'.

Embodiment 2

[0054] Embodiment 2, the application of allele and primer pair

[0055] The experimental materials are several main varieties of Chinese winter wheat, see Table 1 for details.

[0056] Each experimental material was carried out as follows:

[0057] 1. Genomic DNA extraction.

[0058] 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.

[0059] Reaction system for PCR amplification: template DNA about 100ng, ExTaq enzyme (Takara, code: DRR100B) 0.3 μl, GC buffer I 25 μl (Takara, code: DRR20GC I), dNTP (Takara, code: D4030RA, 2.5mM each) 4 μl, upstream The primer and the downstream primer are each 6pmol, and the reaction system is supplemented with sterile ultrapure water to 50μl.

[0060] The reaction program of PCR amplification: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 1 minute, renaturation for 30 seconds (the first renaturat...

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Abstract

The invention discloses a primer pair for assaying germination traits of wheat seed grains and a related molecular marker. The specific primer pair disclosed by the invention consists of a single-stranded DNA fragment shown in a sequence 3 of a sequence table and a single-stranded DNA fragment shown in a sequence 4 of the sequence table. The invention discloses an ordinary wheat gene TaSdr and allelic variation sequences, namely TaSdr-B1a and TaSdr-B1b, of the ordinary wheat gene TaSdr and provides functional markers Sdr2B for assaying the allelic variation of the TaSdr-B1a and the TaSdr-B1b and the relationship between the functional markers and the germination traits of wheat. Through applying the functional markers provided by the invention to molecular-marker-assisted selection, wheat varieties (materials) with higher seed grain dormancy can be quickly screened out, and thus, the pace for breeding new preharvest-sprouting-resistant wheat varieties is accelerated. The primer pair and the related molecular marker have important theoretical significance and economic value in the molecular-marker-assisted selection of the preharvest-sprouting-resistant wheat varieties.

Description

technical field [0001] The invention relates to a pair of primers for identifying germination traits of wheat grains and related molecular markers. Background technique [0002] Pre-harvest sprouting (PHS) refers to the germination of wheat on ears in rainy or humid conditions before harvest. Ear germination leads to a sharp drop in wheat yield and bulk density, and wheat fields with visible ear germination generally reduce yield by about 10%. Ear germination not only affects the yield, but also seriously affects the processing quality of wheat. Even for wheat whose ear germination is not very serious, its processing quality will be greatly affected. The germination of wheat ears significantly reduces the dry and wet gluten content, sedimentation value and rheological properties of dough, making bread or steamed buns sticky and lack of elasticity. During the germination process of wheat ears, the content of amylopectin decreased, the RVA parameters decreased, and the toug...

Claims

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Application Information

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