Functional KASP molecular marker of wheat powdery mildew resistance gene Pm12 and application of functional KASP molecular marker

A powdery mildew resistance gene and molecular marker technology, applied in the fields of biotechnology and genetics and breeding, can solve the problems of difficulty in the development of specific KASP sequences, affecting the efficient utilization of gene Pm12, etc., and achieve high-efficiency breeding and utilization, efficient detection and tracking, high The effect of flux identification

Active Publication Date: 2021-07-13
YANTAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, in the powdery mildew resistance survey of existing popularized varieties and breeding lines in my country, it was found that the proportion of varieties with broad-spectrum resistance among the main popularized varieties in China is very low, less than 5%.
For the gene Pm12, although it has been cloned and has broad-spectrum resistance, because the Pm12 sequence is highly homologous to the three subgroups of common wheat, it is difficult to develop specific KASP sequences, and there is no Pm12 high-pass in the industry. The development and utilization of quantitatively detected KASP molecular markers has seriously affected the efficient use of the gene Pm12 in wheat genetic improvement

Method used

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  • Functional KASP molecular marker of wheat powdery mildew resistance gene Pm12 and application of functional KASP molecular marker
  • Functional KASP molecular marker of wheat powdery mildew resistance gene Pm12 and application of functional KASP molecular marker
  • Functional KASP molecular marker of wheat powdery mildew resistance gene Pm12 and application of functional KASP molecular marker

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Embodiment 1

[0045] Example 1 Obtaining of Wheat Powdery Mildew Resistance Gene Pm12 Functional KASP Molecular Marker KASP-Pm12

[0046] 1. Preparation of DNA amplification template

[0047] (1) DNA amplification template: the transgenic vector carrying Pm12 was constructed and provided by Dr. He Huagang of Jiangsu University, and it is ready for use. The rest of the DNA amplification templates used the CTAB method. The Pm12 transgenic line (provided by Dr. He Huagang of Jiangsu University), the 6SS translocation line 1338-8 carrying Pm12 (provided by Dr. He Huagang of Jiangsu University), the common wheat Chinese spring without Pm12, and the 6VS translocation line wheat carrying Pm21 were extracted respectively. Genomic DNA of leaves at one-leaf stage of variety Yangmai 18 and 6VS translocation line wheat variety Yangmai 22 carrying PmV. The specific operation steps are as follows:

[0048] (1) Cut fresh leaves at the first leaf stage and freeze and grind them with liquid nitrogen. Tak...

Embodiment 2

[0071] Example 2 Application of wheat powdery mildew resistance gene Pm12 functional KASP molecular marker KASP-Pm12 in distinguishing Pm12 from other powdery mildew resistance genes and susceptible materials

[0072] 1. Preparation of DNA amplification template

[0073] The transgenic vector carrying Pm12 was provided by Dr. He Huagang of Jiangsu University, and it was spared. The remaining DNA amplification templates were extracted using the CTAB method. Pm12 transgenic line (provided by Dr. He Huagang, Jiangsu University), 6SS translocation line 1338-8 carrying Pm12 (provided by Dr. He Huagang, Jiangsu University), 6VS translocation line carrying Pm21 wheat variety Yangmai 18, 6VS translocation carrying PmV Wheat variety Yangmai 22, materials carrying known powdery mildew resistance genes Ulka / 8*Cc(Pm2), Tabasco(Pm48), Liangxing 66(PmLX66), D57-5D(PmD57-5D), NCD7(Pm34) , NCD3 (Pm35), Wennong 14 (PmW14), Zhongmai 155 (PmZ155), Jimai 22 (Pm2+Pm52), Yingbo 700 (PmYB), Red Po...

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Abstract

The invention discloses a functional KASP molecular marker of a wheat powdery mildew resistance gene Pm12, the KASP molecular marker is KASP-Pm12, the marker is a nucleotide sequence obtained by a primer amplification DNA template of the KASP-Pm12 designed according to the 3157th basic group of the gene Pm12, and the nucleotide sequence of the KASP molecular marker is as shown in SEQ ID NO: 1; the primer KASP-Pm12 comprises an aegilops spellelsii allelic type upstream primer KASP-Pm12-F, and a wheat allelic type upstream primer KASP-Pm12-H and a shared downstream primer KASP-Pm12-C. The functional KASP molecular marker of the wheat powdery mildew resistance gene Pm12 provided by the invention is applied to identifying whether the Pm12 gene exists in wheat-aegilops spellelsii offspring and wheat varieties/strains or not, and is also applied to assisted selective breeding and pyramiding breeding with other known powdery mildew resistance genes. The invention provides a functional KASP molecular marker capable of rapidly and accurately identifying the gene Pm12 at high throughput, the gene Pm12 in a wheat variety/strain can be efficiently detected and tracked, and efficient breeding utilization of the gene Pm12 is accelerated.

Description

technical field [0001] The invention relates to the fields of biotechnology and genetic breeding, in particular to a functional KASP molecular marker of wheat powdery mildew resistance gene Pm12 and its application. Background technique [0002] Common wheat (Triticum aestivum L., 2n=42, AABBDD), as one of the most important food crops in my country, is very important to ensure national food security and improve people's living standards. However, wheat yield and quality are often severely affected by diseases, especially wheat powdery mildew. As one of the most serious diseases of wheat, wheat powdery mildew is caused by the infection of powdery mildew brucei. Generally, wheat infected with powdery mildew can cause 10-15% of the yield. The annual grain loss caused by wheat powdery mildew in my country is 300,000 tons Above (Chen et al. Positional cloning of PmCH1357 reveals the origin and allelic variation of the Pm2 gene for powdery mildew resistance inwheat. Crop J, 2019,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 刘开昌刘成马朋涛张旭王江春李林志宋文月
Owner YANTAI UNIV
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