Codominant labeling method for identifying wheat polymer glutenin Dx5 and Dx2 subunits

Abstract

The invention discloses a codominant labeling method for identifying wheat polymer glutenin Dx5 and Dx2 subunits. Based on the characteristics of codominant labeling, PCR amplification primers (upstream primer P1: 5'-CCAGCACAAGTGCAACAA-3', downstream primer P2: 5'-TGTCCTAGCTGCAACGGAG -3') are designed, and the PCR amplification primers are used to identify wheat polymer glutenin Dx5 and Dx2 subunits. The method has the advantages of simplicity, stability, easy implementation and capability of accurately identifying Dx5 high-quality subunits, Dx2 subunits, as well as heterozygous types of the two.

Description

technical field [0001] The invention relates to a PCR amplification primer designed based on the characteristics of co-dominant markers and a method for identifying wheat high-scoring glutenin Dx5 and Dx2 subunits, belonging to the field of agricultural biotechnology engineering. Background technique [0002] Wheat storage protein is an important factor to determine the quality of wheat processing, among which gliadin plays a major role in the formation and extensibility of dough, and glutenin is an important factor affecting dough elasticity, which determines the baking quality of bread, especially high molecular weight glutenin The subunit (HMW-Glu) is more closely related to the baking quality of wheat bread, and common wheat varieties usually only contain 3-5 HMW glutenin subunits. The coding gene of HMW-GS is located at the Glu-1 locus of the long arms of the first group of partial homologous chromosomes 1A, 1B, and 1D of common wheat, and they are named Glu-A1, Glu-B1,...

AI Technical Summary

Problems solved by technology

The disadvantages of this method are firstly that it is not precise enough, secondly that the experimental steps are cumbersome and the electrophoresis time is long, which is not conducive to scale identification, and thirdly that the detection and screening work must wait until the next generation of seeds are harvested, which greatly affects the breeding rate

However, this method still has the following disadvantages: first, the flexibility of primer design for allele-specific PCR is not large, and primers can only be designed according to the position of the SNPs site; It is difficult to strictly control under the general environment, which increases the difficulty and cost of specific amplification, because there is only one nucleotide difference between the allele-specific primer and its opposite allele; the third is that the allele-specific PCR marker only It is a dominant marker of the high-quality subunit Dx5, which is a big disadvantage because it cannot distinguish between heterozygous and homozygous genotypes, which brings a lot of inconvenience to MAS breeding work

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