Wheat molecular marker 5668 and application thereof in grain hardness improvement
A technology of wheat grain and molecular markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, food processing, etc., and can solve the problems of moldy core, difficult evaporation of water, and excessive white koji.
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Embodiment 1
[0052] In order to obtain the wheat kernel hardness regulation gene, in conjunction with the prior art, firstly based on the wheat kernel hardness measurement method (single kernel characterization system, SKCS), to the hardness phenotypes of different wheat varieties planted continuously for three years The situation was determined in order to initially determine the differences in wheat grain hardness phenotypes. The specific measurement results are shown in the following Table 1.
[0053] It should be explained that the specific wheat grain hardness measurement method can refer to the following operations:
[0054] After the harvested mature wheat grains were naturally sun-dried, 300 whole seeds without half-grain or insect eyes were selected and poured into the single-grain grain hardness tester SKCS (model is Perten SKCS 4100), run the instrument, and measure the obtained Sample hardness data.
Embodiment 2
[0056] Based on the hardness phenotype data measured in Example 1, two significant loci (1BL: 572152983-573860794) and (1BL: 572638947-572639017bp) were identified and obtained by using the genome-wide association analysis (GWAS) method (partially) See the results figure 1 ), and named the two sites: 5668 and 5668-1, respectively.
[0057] The specific process of obtaining the above-mentioned loci using the genome-wide association analysis (GWAS) method is as follows:
[0058] Genome-wide association studies (GWAS) is a new method for the study of complex traits developed in recent years. In the past few years, many researchers at home and abroad have carried out GWAS on wheat disease resistance, stress resistance, nutritional quality and processing quality. These studies not only greatly enriched the molecular markers available in marker-assisted selection breeding in wheat, but also provided important clues for the exploration and study of the molecular mechanism of these ...
Embodiment 3
[0070] Based on the primer pair design in Example 2, the genotypes of 207 wheat materials were detected, and the correlation between genotypes and hardness phenotypes was identified. The specific process is briefly described as follows.
[0071] First, the genomic DNA of each wheat variety was extracted separately;
[0072] Then, according to the primers designed in Example 2, the DNA extracted above was used as a template, and different wheat samples were subjected to PCR (using BIO-RAD fluorescence quantitative PCR instrument CFX ConnectTM Real-Time System) detection and analysis, and finally determined the 36th base. sequence.
[0073] For specific PCR amplification, the 5 μL amplification system can be designed as follows:
[0074] KASP Mix (2X), 2.5 μL;
[0075] Primer Mix, 0.7 μL;
[0076] MgCl 2 , 0.04μL;
[0077] DNA, 30ng;
[0078] ddH 2 O, supplemented to 5 μL;
[0079] The Primer Mix, per 100 μL:
[0080] 5668-F1, 10uM, 12μL;
[0081] 5668-F2, 10uM, 12μL; ...
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