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A high-throughput detection marker of wheat powdery mildew resistance gene pm2b and its application

A powdery mildew resistance gene and detection marker technology, applied in the fields of biotechnology and genetics and breeding, can solve problems such as loss of resistance of disease resistance genes, and achieve the effect of improving breeding efficiency

Active Publication Date: 2022-04-15
中国科学院遗传与发育生物学研究所农业资源研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the rapid mutation of wheat powdery mildew strains and the widespread use of single disease resistance genes, many disease resistance genes have gradually lost resistance

Method used

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  • A high-throughput detection marker of wheat powdery mildew resistance gene pm2b and its application
  • A high-throughput detection marker of wheat powdery mildew resistance gene pm2b and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: wheat powdery mildew resistance gene Pm2b Development of the KASP molecular marker KASP529

[0028] 1. Materials

[0029] F 1 Backcross twice with Shimai 15 and get BC 2 f 1 There are 188 generation plants.

[0030] 2. Extraction method of wheat genomic DNA

[0031] Genomic DNA of wheat was extracted by CTAB method and stored at -20°C for future use. The specific steps are as follows:

[0032] (1) Take the young leaves of wheat materials, grind them into powder in liquid nitrogen, and put them into 2mL EP tubes (centrifuge tubes);

[0033] (2) Add 600-800 μL CTAB extract to the EP tube, bathe in water at 65°C for 1 hour, and mix by inverting several times during this period;

[0034] (3) Add chloroform equal to the volume of CTAB extract to the EP tube, mix by inverting for 15 min, centrifuge at 12000 rpm for 10 min, and transfer the supernatant to a 1.5 mL EP tube;

[0035] (4) Add 3 times the volume of ice-free ethanol to a 1.5mL EP tube, and settl...

Embodiment 2

[0056] Embodiment 2: wheat powdery mildew resistance gene Pm2b Application of the KASP marker KASP529

[0057] The samples to be tested included 67 wheat materials. The results of gene derivation, molecular marker identification and pedigree tracing showed that 14 of the 67 wheat materials carried known PM2 alleles, the remaining 53 copies do not carry PM2 alleles.

[0058] Extract the DNA of the above-mentioned material as a template, and use the primers of the KASP marker KASP529 developed by the present invention to amplify:

[0059] The upstream sequence is:

[0060] KASP529-FAM (disease resistance locus binding):

[0061] 5'-gaaggtgaccaagttcatgcttgccttggctcagagcatatgtggt-3', as shown in SEQ ID NO.1;

[0062] KASP529-HEX (sensitivity locus binding):

[0063] 5'-gaaggtcggagtcaacggatttgccttggctcagagcatatgtggg-3', as shown in SEQ ID NO.2;

[0064] downstream sequence:

[0065] KASP529-C: 5'-cttaaaatgctttactacctca-3', as shown in SEQ ID NO.3.

[0066] The PCR ampli...

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Abstract

The invention discloses a wheat powdery mildew resistance gene Pm2b High-throughput detection marker and application thereof, the marker is KASP529, and its primers include an upstream primer for a disease-resistant site, an upstream primer for a susceptible site, and a downstream primer; the nucleotide sequence of the upstream primer for a disease-resistant site is as SEQ ID NO.1; the nucleotide sequence of the upstream primer of the susceptible site is shown in SEQ ID NO.2; the nucleotide sequence of the downstream primer is shown in SEQ ID NO.3. KASP marker KASP529 provided by the invention and wheat powdery mildew resistance gene Pm2b Tightly linked, efficient, accurate and high-throughput detection Pm2b Genetic mapping of large populations, applied to Pm2b Fine positioning and map bit cloning; can also achieve PM2 High-throughput detection of other alleles at the locus, for PM2 The in-depth analysis and utilization of loci lay the foundation; the use of this marker for molecular marker-assisted selection can realize breeding in a large population in a short time. PM2 High-throughput screening of disease-resistant genes can effectively improve breeding efficiency and serve wheat disease-resistant molecular breeding.

Description

technical field [0001] The invention relates to the fields of biotechnology and genetic breeding, in particular to a wheat powdery mildew resistance gene Pm2b High-throughput detection markers and their applications. Background technique [0002] wheat( Triticum aestivum L., 2n=6x=42, AABBDD) is one of the most important food crops in the world, and the high and stable yield of wheat plays a vital role in national economic development and national food security. However, in recent years, the poor comprehensive disease resistance of wheat has become the bottleneck restricting the high and stable yield of wheat in my country. Powdery mildew brucella ( Blumeria graminis f. sp. tritici ) caused by wheat powdery mildew has become the main wheat disease in my country, and its occurrence range almost extends to all wheat areas in the country, seriously affecting the yield and quality of wheat. Therefore, the control of wheat powdery mildew has become an urgent need to i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/172C12Q2600/156C12Q2600/13C12Q2531/113C12Q2563/107
Inventor 安调过靳玉丽韩帼豪麻菲菲生媛王婧马朋涛许红星
Owner 中国科学院遗传与发育生物学研究所农业资源研究中心