Development and application of KASP molecular marker of rice blast resistance gene Pid3-A4

A rice blast disease and molecular marker technology, applied in the field of inductor rice breeding, can solve the problems of low detection efficiency, unsuitable high-throughput molecular detection platform, and aerosol pollution of the environment, so as to reduce pollution and reduce field planting. Scale, the effect of reducing breeding costs

Pending Publication Date: 2021-10-22
华美瑞康北京国际生物技术研究院有限公司
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  • Description
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Problems solved by technology

Among them, although SSR markers, InDel markers or enzyme-cut markers can be used for molecular marker-assisted selection, the detection efficiency is low, and aerosol pollution may be generated at the same time, which is not suitable for high-throughput molecular detection platforms

Method used

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  • Development and application of KASP molecular marker of rice blast resistance gene Pid3-A4
  • Development and application of KASP molecular marker of rice blast resistance gene Pid3-A4
  • Development and application of KASP molecular marker of rice blast resistance gene Pid3-A4

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1: Development of KASP marker closely linked to rice blast resistance gene Pid3-A4

[0040] The development process of the KASP marker closely linked to the rice blast resistance gene Pid3-A4 is as follows: figure 1shown. According to literature reports, a SNP locus [CA / AG] closely linked to the Pid3-A4 gene was obtained (the physical location of the rice reference genome is chromosome 6 10389855-10389856bp).

[0041] 1. Determination of primers:

[0042] The tightly linked SNP locus of Pid3-A4 gene was anchored at the 13058850bp position of rice chromosome 6 (Huang Weiheng, Huang Zhiyuan, Tang Li, et al. Development and application of specific InDel molecular markers for rice blast resistance Pid3 / Pid3-A4 gene[J] . Hybrid Rice, 2020(2):68-74.). The flanking sequence of closely linked SNP sites of Pid3-A4 gene was downloaded from NCBI database, and three sets of KASP primers were designed using Primer5.0 software. After detection by the ArrayTape platform of...

Embodiment 2

[0070] Example 2: Application of KASP marker closely linked to rice blast resistance gene Pid3-A4 in molecular marker-assisted selection of rice blast resistance plants

[0071] see image 3 , in order to detect the practicability of the KASP-A4 primer of the present invention, the F1 population was obtained by crossing the cultivated rice blast-resistant material 311100 with the rice blast-susceptible material Daohuaxiang No. 2, and 90 F2 natural segregating populations were produced by natural selfing of F1. For the segregating population Carry out KASP marker detection and disease-resistant phenotype verification (Table 2), the implementation method of marker detection and disease-resistant phenotype verification refers to Example 1. Through phenotypic and genotypic detection and analysis of segregating populations (e.g. image 3 shown), among the 90 isolated populations, only 6 genotypes were inconsistent with the phenotype results, and the consistency result between the ...

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Abstract

The invention provides development and application of a KASP molecular marker closely linked with a rice blast resistance gene Pid3-A4. Rapid gene typing identification is carried out on an SNP site closely linked with the rice blast resistance gene Pid3-A4 by utilizing a KASP technology, and the KASP molecular marker can be applied to commercial molecular breeding with high, medium and low throughput; meanwhile, the phenotype selection efficiency of the molecular marker KASP-A4 reaches 93.33%, and the rice blast resistance gene Pid3-A4 can be rapidly and accurately detected in different germplasm resources of rice. In addition, tedious procedures such as enzyme digestion, electrophoresis and sequencing are not needed in the detection process, aerosol pollution and use of toxicants such as ethidium bromide EB are reduced, molecular marker-assisted selection can be carried out in the early breeding stage, the field planting scale of breeding populations is reduced, the breeding cost is reduced, and the breeding process is accelerated.

Description

technical field [0001] The invention relates to the technical field of inductor rice breeding, in particular to the development and application of a KASP molecular marker of rice blast resistance gene Pid3-A4. Background technique [0002] Rice is an important crop second only to wheat in terms of cultivated area and total output in the world. The cultivated area of ​​rice in my country is second only to India, but the total rice output ranks first among the rice-producing countries in the world. The fruit of rice is rice, which can be used as food, and can also be used as industrial raw materials for brewing wine and sugar. Mats, grass paper, etc., and the rice stalks can also be used to make boards and fertilizers. [0003] Blast has another name called rice fever, burning blast, knocking head blast, etc. It is one of the most serious diseases of rice, and the rice lost due to rice blast is 10% to 30% of its annual output (Skamnioti P, Gurr S J. Against the grain:safegu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11C40B40/06
CPCC12Q1/6895C12Q1/6858C40B40/06C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113C12Q2563/107
Inventor 马江涛颜光华
Owner 华美瑞康北京国际生物技术研究院有限公司
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