Primer pair for assaying thousand-grain weight of wheat and related molecular marker

A thousand-grain weight, specific primer pair technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of poor comparability, poor repeatability, and small contribution rate of grain weight and its constituent elements to the phenotype. and other problems to achieve the effect of accelerating the pace of breeding

Inactive Publication Date: 2013-04-24
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

Due to the influence of mapping population, genetic background, QTL mapping method, marker type and other factors, the results of different researchers are not comparable
In addition, since the phenotypic contribution rate of most QTLs to grain weight and its components is small, and the reproducibility is poor in different years and environments, it still cannot meet the needs of molecular marker-assisted selection.

Method used

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  • Primer pair for assaying thousand-grain weight of wheat and related molecular marker
  • Primer pair for assaying thousand-grain weight of wheat and related molecular marker
  • Primer pair for assaying thousand-grain weight of wheat and related molecular marker

Examples

Experimental program
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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 TaGS gene, two allelic variant forms of the TaGS gene in wheat, one as shown in sequence 1 of the sequence table (named allele TaGS-D1a ), the other is shown in sequence 2 of the sequence listing (named TaGS-D1b). The comparison spectrum of the two allelic variant forms of the TaGS gene is shown in figure 1 , the start codon and stop codon are shown in boxes, the underlined part is an exon, the shaded part is a sequence polymorphic site, and the missing part of the sequence is replaced by a short horizontal line.

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

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

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

Embodiment 2

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

[0055] The experimental materials are 106 Chinese winter wheat main varieties, 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, rTaq enzyme (Takara, code: DX100B) 0.2 μl, GC buffer Ⅰ 10 μl (Takara, code: DRR20GC I), dNTP (Takara, code: D4030RA, 2.5mM each) 2 μl, upstream The primer and the downstream primer are each 6pmol, and the reaction system is supplemented with sterile ultrapure water to 20μl.

[0060] The reaction program of PCR amplification: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min, annealing at 52°C for 1 min, extension at 72°C for 1 min, a tota...

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Abstract

The invention discloses a primer pair for assaying the thousand-grain weight of wheat 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 allelic variation sequences, namely TaGS-D1a and TaGS-D1b, of an ordinary wheat gene TaGS and provides functional markers for assaying the allelic variation of the TaGS-D1a and the TaGS-D1b and the relationship between the functional markers and the thousand-grain weight of the wheat. Through applying the functional markers provided by the invention to molecular-marker-assisted selection, wheat varieties (materials) with higher thousand-grain weight can be quickly screened out, and thus, the pace for breeding new high-yielding 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 high-yielding wheat varieties.

Description

technical field [0001] The invention relates to a pair of primers for identifying thousand-grain weight of wheat and related molecular markers. Background technique [0002] Wheat is an important food crop in my country. The high and stable yield of wheat is an important guarantee for national food security. The level of its output directly affects the living standards of our people and national food security. With the increase of population, the reduction of arable land and the continuous increase of grain production cost, high-yield breeding will be the eternal theme of wheat breeding in my country. Thousand-grain weight is directly related to wheat yield. Therefore, it is of great significance for high-yield breeding in my country to study the genetic basis of genes controlling wheat grain size, clone grain-weight-related genes, discover excellent allelic variations and develop their functional markers by means of modern molecular biology. . [0003] The components of yi...

Claims

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Application Information

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Patent Type & Authority Applications(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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