SNP sites remarkably associated with wheat powdery mildew resistance and application thereof in genetic breeding

A technology of wheat powdery mildew and loci, which is applied in the application field of genetics and molecular breeding, can solve the problems of high cost of deep sequencing, difficulty for breeders to master and use, and achieve the effect of low accuracy

Inactive Publication Date: 2021-03-16
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared to the huge genome (16G) of wheat, the cost of deep sequencing is still high, and GBS data processing, sequence comparison, genotyping and other processes have high requir

Method used

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  • SNP sites remarkably associated with wheat powdery mildew resistance and application thereof in genetic breeding
  • SNP sites remarkably associated with wheat powdery mildew resistance and application thereof in genetic breeding
  • SNP sites remarkably associated with wheat powdery mildew resistance and application thereof in genetic breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Identification of SNPs by Simplified Genome Sequencing

[0036] 1.1 Sequencing materials

[0037] 768 wheat cultivars (lines) were selected for simplified genome sequencing to identify SNPs. These wheat varieties (lines) are mainly from the main wheat producing areas in my country, including the Huanghuai wheat area, the northern winter wheat area, the middle and lower reaches of the Yangtze River wheat area and the southwest wheat area.

[0038] Research methods

[0039] 1.2.1 Extraction of wheat genomic DNA

[0040]Seedling leaves were used to provide genomic DNA. DNA was extracted using the improved CTAB method. Specific operation: Take young wheat leaves in a 2mL centrifuge tube, freeze them with liquid nitrogen and grind them into powder on a tissue grinder; (b) add 800 μL CTAB to the 2mL tube, place in a 65°C water bath for 90 minutes, and gently Shake 5-8 times to fully lyse the DNA; (c) add 800 μL chloroform isoamyl alcohol (volume ratio 24:1) and...

Embodiment 2

[0049] Example 2 Genome-wide Identification of Powdery Mildew Resistance Gene Loci

[0050] 2.1 Materials

[0051] 768 accessions of wheat varieties (lines) from the main wheat producing areas in my country, including the Huanghuai wheat area, the northern winter wheat area, the middle and lower reaches of the Yangtze River wheat area, and the southwest wheat area, are the same as in Example 1.

[0052] 2.2 Method

[0053] 2.2.1 Identification of powdery mildew resistance

[0054] In order to identify the resistance of materials to powdery mildew, these 768 wheat varieties (lines) were planted in the experimental station of Shandong Agricultural University in Tai'an in 2017-2018 (TA17), 2018-2019 (TA18), and 2019-2020 (TA19) The disease identification nursery is used for identification and identification of powdery mildew resistance by artificial inoculation. At the same time, these materials were also planted in Yantai (YT), Luoyang (LY) and Guiyang (GY) in 2017-2018, and ...

Embodiment 3

[0068] Example 3 Wheat Powdery Mildew Resistance Gene qPm6A.3 verification and mapping

[0069] 3.1 Materials

[0070] The research materials include the powdery mildew-resistant parent 2013BP24, the susceptible parent Xumai 32 and the F 6 There are 126 recombinant inbred lines in total in the group of recombinant inbred lines. Huixianhong and Mingxian 169 were used as susceptible controls.

[0071] 3.2 Method

[0072] 3.2.1 Phenotype identification of disease resistance

[0073] With embodiment 1, the identification method of powdery mildew resistance

[0074] 3.2.2 DNA extraction

[0075] With embodiment 1.

[0076] 3.2.3 KASP primer design and dilution:

[0077]According to the results of GWAS analysis, the powdery mildew resistance QTL loci qPm6A.3 According to the significantly associated SNP (6A_86486561), a set (a total of three) of KASP primers k6A86486 was designed according to the SNP site information. Dilute the three primers of this set of primers to 10...

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Abstract

The invention discloses a group (58 wheat varieties) of SNP sites remarkably associated with wheat powdery mildew resistance and an application method thereof in genetic breeding. The SNP sites are identified by performing simplified genome sequencing (GBS) on 768 wheat varieties and excellent strains to explore SNP sites and then performing disease resistance identification and whole genome association analysis. The group of SNP sites are high in accuracy, can be converted into KASP markers and SNP chips, and are used for positioning, fine mapping and candidate gene identification of wheat powdery mildew resistance genes and wheat powdery mildew resistance whole genome selective breeding.

Description

technical field [0001] The invention relates to the technical field of plant molecular markers, in particular to a group of SNPs significantly associated with wheat powdery mildew resistance gene loci and their application in genetic and molecular breeding of powdery mildew resistance. Background technique [0002] Wheat powdery mildew is caused by powdery mildew Blumeria graminis f. sp. tritici ) caused by an airborne fungal disease, is a worldwide disease, are distributed in major wheat-producing countries. In recent years, with the adjustment of the farming system and the continuous change of the climate, the disease has become increasingly serious in several major wheat regions such as Northeast my country, North China, Huanghuai and Southwest China. The disease mainly occurs at the seedling stage and the adult plant stage, and mainly damages the leaves and leaf sheaths. When the disease is severe, the glumes and awns can also be damaged. Generally, wheat powdery mi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2600/13C12Q2531/113C12Q2537/161Y02A50/30
Inventor 刘树兵庞昀龙张绘蕊李文辉董磊武玉叶
Owner SHANDONG AGRICULTURAL UNIVERSITY
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