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Rice bacterial blight resistance gene xa5 specific molecular marker and application thereof

A technology of bacterial blight resistance and molecular markers, which is applied in rice bacterial blight resistance gene xa5 specific molecular markers and its application field, can solve the problems of low breeding efficiency and inaccurate results of disease resistance identification, etc. Breeding cycle, high accuracy, and cost-saving effects

Inactive Publication Date: 2019-01-15
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In conventional disease resistance breeding, inaccurate identification of disease resistance results in low breeding efficiency

Method used

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  • Rice bacterial blight resistance gene xa5 specific molecular marker and application thereof
  • Rice bacterial blight resistance gene xa5 specific molecular marker and application thereof
  • Rice bacterial blight resistance gene xa5 specific molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Development of specific molecular markers for rice bacterial blight resistance gene xa5

[0026] (1) Test materials:

[0027] xa5 gene donor parent: IRRBB5;

[0028] Recipient parents: Huhan 1B, Huhan 7B, Hanhui 3, Hanhui 37, Hanhui 49, Huazhan, Huanghuazhan;

[0029] (2) Extraction of rice genomic DNA (for rice genomic DNA methods, please refer to (Lou Qiaojun, Chen Liang, Luo Lijun. Comparison of three rapid extraction methods of rice genomic DNA[J]. Molecular Plant Breeding, 2006, 3(5): 749-752));

[0030] (3) Development of xa5 gene-specific molecular markers:

[0031] Such as figure 1 , detection of rice bacterial blight resistance gene xa5 by four-primer amplification hindered mutation system PCR method: four-primer ARMS-PCR is a special method for identifying single nucleotide polymorphisms in DNA developed on the basis of ordinary PCR technology The basic principle of the method is that Taq DNA polymerase lacks 3'→5' exonuclease activity, and can...

Embodiment 2

[0040] Example 2, detection of parental polymorphism of rice bacterial blight resistance gene xa5 specific molecular marker:

[0041] (1) Test materials:

[0042] xa5 gene donor parent: IRRBB5;

[0043] Recipient parents: Huhan 1B, Huhan 7B, Hanhui 3, Hanhui 37, Hanhui 49, Huazhan, Huanghuazhan;

[0044](2) Using four specific molecular markers xa5-OUT F, xa5-OUT R, xa5-IN F and xa5-IN R to perform PCR amplification on the rice donor and recipient parent material, the PCR amplification system is: 20ng rice genome DNA template, 10 uL Taq PCRMastermix (Tiangen Biochemical Technology (Beijing) Co., Ltd.), 0.5 uL each of the four primers at 10 uM, supplemented with dd H2O to 20uL; 30 s, 30 s at 55°C, 1 min at 72°C for 30 cycles, and a final extension at 72°C for 5 min;

[0045] (3) Take 8 uL of the PCR product and put it on 2% agarose gel electrophoresis for detection. The results are as follows: figure 2 As shown, the 486bp band amplified by the outer primers xa5-OUT F and x...

Embodiment 3

[0046] Example 3: Verification and application of rice bacterial blight resistance gene xa5 specific molecular marker

[0047] In order to verify that the mutation site identified by this marker is a single-gene segregation in the segregation population, the above-mentioned specific molecular markers were used to amplify the DNA of 96 individual plants of the BC1F2 population constructed by Hanhui 3 and IRBB5, and the results are as follows: image 3 As shown, the electrophoresis products present three types of bands, that is, the characteristic bands of 486bp and 390bp for the homozygous xa5 gene; the characteristic bands of 486bp, 390bp and 143bp for the heterozygous xa5 gene; and a characteristic band of 143bp.

[0048] Statistics found that the segregation ratio of the three genotypes was 23:54:19, which was consistent with the Mendelian single-gene segregation ratio of 1:2:1 by chi-square test (χ2=1.172 0.05 =5.99), through artificial inoculation and identification of X...

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Abstract

The invention discloses the development and the application of a rice bacterial blight resistance gene xa5 specific molecular marker. Four molecular marker primers were designed and added to the samePCR reaction system to amplify different rice genomic DNA. If there are two characteristic bands of 486bp and 390bp, it indicates that the sample is homozygous for xa5 gene. If there are two characteristic bands of 486 bp and 143 bp, the sample does not contain xa5 gene; If three characteristic bands of 486 bp, 390 bp and 143 bp are simultaneously present, it indicates that the sample is xa5 geneheterozygous. The four-primer PCR molecular marker is a co-dominant marker, which can effectively distinguish three different genotypes of rice bacterial blight resistance gene xa5, improve the selection efficiency of the gene, and accelerate the improvement and breeding process of rice bacterial blight resistance.

Description

technical field [0001] The invention relates to a biomolecular marker and its application technology, in particular to a rice bacterial blight resistance gene xa5 specific molecular marker and its application. Background technique [0002] Bacterial blight is one of the main diseases of rice, which leads to a large reduction and instability of rice yield, and also affects rice quality. Practice has proved that cultivating and planting disease-resistant varieties is one of the most cost-effective methods to control this disease, and the key to cultivating disease-resistant varieties lies in excavating and utilizing excellent disease-resistant gene resources. So far, 42 bacterial blight resistance genes have been identified from different rice sources. xa5 is a cloned recessive gene for resistance to bacterial blight. There are two base differences (A116T and G117C) in the xa5 gene sequence between the disease-resistant variety IRBB5 and the susceptible variety Nipponbare, re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘毅刘国兰王加红张安宁王飞名孔德艳毕俊国余新桥罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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