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Molecular marker for identifying peroxidase activity of wheat grains and application of molecular marker

A technology of wheat and activity, which is applied in the field of molecular markers to identify wheat grain peroxidase activity, and can solve the problems that have not yet been studied in depth

Active Publication Date: 2020-10-02
XINJIANG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Shi Jia et al. used two natural populations combined with the 90K iSelect chip to perform GWAS on wheat grain POD activity for the first time. After finding the 2A, 2D, 7B and 7D stable genetic loci, they cloned and developed the 7D chromosome with the largest locus contribution rate. The POD activity gene TaPod-D1 functional markers POD-7D1 and POD-7D6 have been reported to clone the main genes involved in POD biosynthesis in higher plants, but their regulatory mechanism has not been studied in depth. In order to more accurately identify the POD activity of wheat varieties Genotype, it is necessary to clone the important main effect site gene, and develop co-segregated functional markers according to the sequence of different alleles of the gene site, further improve the accuracy of molecular marker selection and breeding efficiency, and provide a basis for improving wheat POD activity. Effective molecular markers will eventually lay the foundation for molecular marker-assisted breeding and map-based cloning of the gene

Method used

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  • Molecular marker for identifying peroxidase activity of wheat grains and application of molecular marker
  • Molecular marker for identifying peroxidase activity of wheat grains and application of molecular marker
  • Molecular marker for identifying peroxidase activity of wheat grains and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Embodiment 1, the establishment of molecular markers and methods for identifying wheat grain peroxidase (Peroxidase, POD) activity

[0108] 1. Molecular markers for identification of wheat grain peroxidase activity

[0109] There are 3 allelic variant forms of TaPod-A1 gene in wheat, the corresponding alleles are TaPod-A3a, TaPod-A3b and TaPod-A3c, and the nucleotide sequence of the allele TaPod-A3a is sequence 7; etc. The nucleotide sequence of allele TaPod-A3b is sequence 8; the nucleotide sequence of allele TaPod-A3c is sequence 9.

[0110] Based on the three allelic variants of the TaPod-A1 gene, specific primer pair 1 (to amplify a partial fragment of the allele TaPod-A3a), primer pair 2 (to amplify a partial fragment of the allele TaPod-A3b) and primer pair 3 ( Amplified allele TaPod-A3c partial fragment) as follows:

[0111] The upstream primer of primer pair 1 (sequence 1): 5'-CACGAGACGCTGTGGAAGGACAG-3';

[0112] The downstream primer of primer pair 1 (sequen...

Embodiment 2

[0149] Embodiment 2, molecular marker identification wheat grain peroxidase activity

[0150] Experimental materials: 25 Chinese winter wheat main varieties (as shown in Table 1), the public can obtain from the National Germplasm Bank of the Institute of Crop Science, Chinese Academy of Agricultural Sciences.

[0151] 1. Detection of POD activity in wheat grains

[0152] The wheat varieties shown in Table 1 were planted in Manas, Xinjiang, and the POD activity of wheat grains was tested after harvesting. The POD activity of each wheat material was tested twice. If the error of the two test results exceeds 10%, it is necessary to carry out The test was repeated for the third time, and the results were averaged.

[0153] The method for measuring the POD activity of the grain is as follows:

[0154] 1. Preparation of whole wheat flour

[0155] (1) Weigh 5 g of wheat grains with impurities removed and grind them with a cyclone mill (LaboratoryMill 120, Sweden) equipped with a 0...

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Abstract

The invention discloses a molecular marker for identifying the peroxidase activity of wheat grains and an application of the molecular marker. The invention provides a complete set of primers for identifying the peroxidase activity of wheat. The complete set of primers consists of a primer pair 1, a primer pair 2 and a primer pair 3, wherein the primer pair 1 is composed of single-stranded DNA asshown in a sequence 1 and a sequence 2; the primer pair 2 is composed of single-stranded DNA as shown in a sequence 3 and a sequence 4; and the primer pair 3 is composed of single-stranded DNA as shown in a sequence 5 and a sequence 6. The molecular marker disclosed by the invention can be used for rapidly identifying allelic variation of TaPod-A3a, TaPod-A3b and TaPod-A3c and a correlation relationship between the allelic variation and the POD activity of the wheat, and screening out a wheat variety with relatively high POD activity, so that the breeding pace of a new high-quality wheat variety is accelerated, and the molecular marker has important theoretical significance and economic value in molecular marker-assisted selection of wheat varieties with high POD activity.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular marker for identifying wheat grain peroxidase activity and an application thereof. Background technique [0002] Peroxidase (POD) in wheat grains has browning and bleaching effects on flour and its products. The higher the activity of POD, the whiter the color of wheat flour. With the improvement of people's living standards, the quality of wheat has become an important goal of wheat breeding and production in my country, and it is also an important factor affecting the competitiveness of wheat industries. Wheat quality mainly includes milling quality, processing quality, nutritional quality and starch quality. Wheat milling quality refers to the quality characteristics of wheat during the milling process, which is mainly determined by thousand-grain weight, bulk density, ash content and flour extraction rate. Processing quality is mainly determined by glute...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 耿洪伟夏先春严勇亮王丽丽王继庆战帅帅任毅
Owner XINJIANG AGRI UNIV
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