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Chain KASP molecular markers, primers and application of wheat anti-yellow-rust gene QYr.nwafu-6BL.2

A technology of qyr.nwafu-6bl.2 and molecular markers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of difficult to overcome resistance, loss of disease resistance of varieties, wheat bar The problem of single source of resistance to rust disease can shorten the time

Active Publication Date: 2019-05-10
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the outstanding problems faced by the utilization of wheat disease-resistant varieties in my country are: the resistance source of wheat stripe rust is relatively single and scarce; in production, varieties with a single resistance gene are planted in a large area, so that new toxic races quickly develop into Dominant race, leading to "loss" of variety disease resistance
Mildly effective disease resistance genes usually have some changes in the way they act on the pathogen, so it is more difficult for the pathogen to overcome its resistance

Method used

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  • Chain KASP molecular markers, primers and application of wheat anti-yellow-rust gene QYr.nwafu-6BL.2
  • Chain KASP molecular markers, primers and application of wheat anti-yellow-rust gene QYr.nwafu-6BL.2
  • Chain KASP molecular markers, primers and application of wheat anti-yellow-rust gene QYr.nwafu-6BL.2

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A linked KASP molecular marker of wheat stripe rust resistance gene QYr.nwafu-6BL.2 is provided, the molecular markers are molecular marker KASP-630 and molecular marker KASP-162;

[0040] Among them, the nucleotide sequence of the molecular marker KASP-630 is shown in SEQ ID NO.1, which is: 5'-gagcgagtgaccgagcgtacacttacaaatgatga[g / t]ggccgagttactggtgagacgtaggcatatccttg-3', and the nucleotide sequence is the 36th base from the 5' end is the SNP site;

[0041] The nucleotide sequence of the molecular marker KASP-162 is shown in SEQ ID NO.2, which is: 5'-cattttgcttttccaaagttgacggatcgttgaaa[a / g]ttgtggattggaaagatgaaagatccacgatccgt-3', and the 36th base from the 5' end of the nucleic acid sequence is SNP site.

[0042] A kind of PCR-specific amplification primer of the linked KASP molecular marker of the above-mentioned wheat stripe rust resistance gene QYr.nwafu-6BL.2, the primer of molecular marker KASP-630 (synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.) includ...

Embodiment 2

[0065] 1. Genetic population construction and genetic analysis

[0066] (1) Wheat material for testing: CIMMYT germplasm P10078 (Triticum aestivum) has good adult plant resistance to stripe rust, and this material is recorded in the following references: CIMMYT (1983) Report on wheat improvement. International Maize and Wheat Improvement Center, Mexico, p 20. The wheat variety "Mingxian 169" susceptible to stripe rust was bred by Shanxi Mingxian School (the predecessor of Shanxi Agricultural University) in the 1920s and can be obtained from the National Crop Germplasm Bank. F 1 , F 1 F 2 , F 2 Continuous selfing of a single plant uses the method of "single seed transmission" to obtain a recombinant inbred line (recombinant inbredline, RIL) population, which is used for genetic analysis and QTL mapping. Common wheat Xiaoyan 22 is a cultivar that can be obtained from the National Crop Germplasm Bank.

[0067] (2) Phenotype evaluation and phenotype data analysis of the test ...

Embodiment 3

[0072] Feasibility verification of molecular markers KASP-630 and KASP-162 and their special primer sets for stripe rust resistance genes:

[0073] Method: KASP-630 and KASP-162 primer sets were used to detect stripe rust resistance genes in 176 wheat cultivars.

[0074] 1. Detect 176 samples of species to be tested by using the method of the embodiment with the KASP-630 primer set, and the template solution with a DNA concentration of 100 ng / μl.

[0075] If the genotype of the target SNP in the PCR amplification product is non-CC homozygous, the wheat to be tested is a candidate wheat with stripe rust resistance traits; if the genotype of the target SNP in the PCR amplification product is CC homozygous, Then the wheat to be tested is a candidate wheat with stripe rust traits; the non-CC homozygous type is AA homozygous or AC heterozygous, and the results are as follows: Figure 4 .

[0076] Detect 176 species to be tested with KASP-162 primer set:

[0077] 2. Use the KASP-...

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Abstract

The invention belongs to the technical field of wheat gene detection, and particularly relates to chain KASP molecular markers, primers and application of a wheat anti-yellow-rust main-effect site gene QYr.nwafu-6BL.2. The molecular markers include the molecular marker KASP-630 and the molecular marker KASP-162. By using the two molecular markers in cooperation, the prediction accuracy of the yellow rust main-effect site QYr.nwafu-6BL.2 is up to 98%.

Description

technical field [0001] The invention belongs to the technical field of wheat gene detection, in particular to a linked KASP molecular marker, primer and application of wheat stripe rust resistance gene QYr.nwafu-6BL.2. Background technique [0002] Wheat is the most important food crop in my country. Ensuring high and stable wheat yields is not only a necessity for the basic needs of people's lives, but also a strategic choice for maintaining national and world food security. There are many factors that affect the high and stable yield of wheat. The environment can be roughly divided into biotic stress and abiotic stress. Fungal diseases are important biotic stress factors that affect the safety of wheat production. Wheat stripe rust is a worldwide wheat disease, which occurs in most wheat producing areas in the world. China is one of the countries with the largest incidence of wheat stripe rust and the most serious losses. Wheat stripe rust is an airborne leaf disease caus...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
Inventor 吴建辉曾庆东刘胜杰韩德俊康振生
Owner NORTHWEST A & F UNIV
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