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KASP functional molecular marker of rice blast resistance gene Pi2 and application of KASP functional molecular marker

A technology of rice blast and functional molecules, which is applied in the field of rice breeding, can solve problems such as difficult to achieve batch and accurate detection, complicated molecular marker operations, and obstacles to large-scale applications, so as to achieve good commercial application prospects, improve the efficiency of variety breeding, and detect The results are accurate and reliable

Inactive Publication Date: 2019-02-26
RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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  • Application Information

AI Technical Summary

Problems solved by technology

Pi2 is a gene with broad-spectrum resistance to rice blast physiological races, which has very important application value for rice blast resistance breeding, but the molecular markers developed for the molecular marker-assisted selection breeding of this gene are complicated. , it is difficult to achieve batch and accurate detection, which seriously hinders the large-scale application of this gene in the process of rice MAS breeding
And the EB or polyacrylamide used in the detection process of these marks is easy to pollute the environment and cause harm to the human body

Method used

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  • KASP functional molecular marker of rice blast resistance gene Pi2 and application of KASP functional molecular marker
  • KASP functional molecular marker of rice blast resistance gene Pi2 and application of KASP functional molecular marker
  • KASP functional molecular marker of rice blast resistance gene Pi2 and application of KASP functional molecular marker

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Experimental program
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Effect test

Embodiment 1

[0051] The primer development preparation method of molecular marker W-Pi2, its steps are:

[0052] According to existing literature reports, there is a nucleotide variation GCAGGA / GTG TTA on the 787th and 788th codons of the Pi2 gene. The present invention finds that the rice blast resistance variety Huang Guangyouzhan and the rice blast resistance variety Guang There is a polymorphic difference at this site in Lu Ai 4, Huang Guangyouzhan is a GCA GGA genotype, and Guang Lu Ai 4 is a GTG TTA genotype, so the 100bp sequences on the left and right sides of this site were extracted to develop markers W-Pi2, the marker sequence is shown in SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3.

[0053] 1) Extract the DNA of Huangguangyouzhan and Guangluai 4 as standard DNA samples of the resistant allele type and the susceptible allele type, and mix equal amounts of DNA from the two varieties as the heterozygous genotype standard DNA samples.

[0054] 2) Apply the method of the molecular ma...

Embodiment 2

[0057] Molecular marker W-Pi2 identifies the allele type of rice blast resistance gene in rice varieties, and the steps are:

[0058] 1) extracting the DNA of 40 rice varieties to be tested;

[0059] 2) Apply the method of the molecular marker and genotyping provided by the content of the invention, carry out KASP detection to 40 rice varieties to be tested and 4 standard sample DNAs in Example 1, and the detection results are as follows figure 2 shown.

[0060] 3) The X-axis is the coordinate axis of the HEX fluorescence signal, the Y-axis is the coordinate axis of the FAM fluorescence signal, the small square is the distribution point of the resistant allele type, the small triangle is the distribution point of the heterozygous genotype, and the small circle is the susceptible one, etc. The genotype distribution points, and the small diamonds are the distribution points of the negative control. The genotyping results showed that the marker genotypes of 28 varieties were C...

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Abstract

The invention discloses a KASP functional molecular marker of a rice blast resistance gene Pi2 and an application of the KASP functional molecular marker, and relates to the technical field of rice breeding. The KASP functional molecular marker is marked as variant GCA GGA / GTG TTA on codons in 787-poisition and 788-poisition of the gene Pi2. With the adoption of the KASP functional molecular marker as well as primers and a kit thereof, an allelotype of the rice blast resistance gene Pi2 of a breeding material can be detected in the early stage (a seed or seedling stage), rice blast resistanceof the breeding material is predicted, and the breeding material is accurately screened and is not required to be planted in a disease nursery for evaluation. Therefore, genetic improvement of a ricevariety with rice blast resistance is promoted, and efficiency of variety breeding is improved. An SNP (single nucleotide polymorphism) site is detected with a KASP method, the detection method is simple to operate and low in cost, a detection result is accurate and reliable, and the KASP functional molecular marker is environmentally friendly, harmless to a human body and suitable for high-throughput large-scale commercial breeding application.

Description

technical field [0001] The invention relates to the technical field of rice breeding, in particular to a KASP functional molecular marker of rice blast resistance gene Pi2 and an application thereof. Background technique [0002] Rice blast is a worldwide rice disease, which can cause 10-30% yield reduction every year. Breeding and planting of disease-resistant varieties is the safest and most effective way to control rice blast. However, the traditional breeding method of hybridization combined with phenotype often needs to increase the backcross population because of the inaccurate grasp of the phenotype at the macro level, which greatly increases the workload and cost of breeding. [0003] Molecular marker-assisted selection (MAS) can track target traits on a genetic basis, and select individual plants containing the target gene for hybridization (backcrossing), which can not only accurately carry out breeding for target traits, but also reduce The size of the backcross...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858C12Q1/6895C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113
Inventor 王重荣周少川李宏黄道强周德贵王志东陈宜波吴玉坤赵雷龚蓉潘阳阳杨义强
Owner RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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