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KASP molecular marker for detecting rice blast resistance broad-spectrum gene Pil and application

A molecular marker and rice blast resistance technology, applied in the field of molecular biology, can solve problems such as being easily restricted by environmental conditions, easy to cause errors in identification results, and low selection efficiency, and achieve accurate and reliable detection results, convenient operation, and high resolution efficiency low effect

Active Publication Date: 2019-10-25
广西马坪壮欢科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional rice resistance breeding is phenotypic selection of plants through resistance identification, which takes a long time and is easily limited by environmental conditions. The identification results are prone to errors and the selection efficiency is low.

Method used

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  • KASP molecular marker for detecting rice blast resistance broad-spectrum gene Pil and application
  • KASP molecular marker for detecting rice blast resistance broad-spectrum gene Pil and application
  • KASP molecular marker for detecting rice blast resistance broad-spectrum gene Pil and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Development of Molecular Marker of Rice Blast Resistance Broad Spectrum Gene Pi1

[0031] The present invention performs whole-genome sequencing on six rice varieties (N213, N91, N58, N1209, N1208, and N1298) selected and bred by the Rice Research Institute of the Academy of Agricultural Sciences of Guangxi Zhuang Autonomous Region, and blasts the sequencing results with the rice genome published on NCBI. Analysis, by comparing with the sequenced Pi1 gene sequence, the original SNP site was obtained, and then the original SNP site was verified experimentally, and two SNP molecules with good co-segregation with resistant varieties and KASP reaction amplification effect were selected Mark the site.

[0032] The above two SNP molecular marker sites are:

[0033] SNP site 1: located at base 30654770 of rice chromosome 11, the molecular marker polymorphism is A / T, the sequence of 401bp before and after the SNP site 1 is shown in SEQ.ID.NO.7, the 201bp The base is...

Embodiment 2

[0038] Example 2 Application of KASP Molecular Marker of Rice Blast Resistance Broad Spectrum Gene Pi1

[0039] (1) Extract the rice genomic DNA to be tested

[0040] A total of 94 rice samples to be tested were extracted from genomic DNA, and the quality and concentration of the extracted rice genomic DNA were detected using 1% agarose gel electrophoresis and SMA4000 to ensure that the genomic DNA bands were complete without tailing, and that A260 / 280 is between 1.8 and 2.0, and A260 / 230 is between 1.8 and 2.0.

[0041] (2) KASP specific PCR reaction

[0042] The KASP reaction was performed on the genomic DNA of the rice material to be tested. The KASP reaction was carried out on the LGC SNPline genotyping platform, and the LGC SNPline genotyping platform and experimental reagents were purchased from Fluorescent LGC Company.

[0043] The PCR reaction system and conditions for KASP detection are as follows:

[0044] Table 1 PCR reaction system for KASP detection

[0045] ...

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Abstract

The invention discloses a KASP molecular marker for detecting a rice blast resistance broad-spectrum gene Pil and application. The molecular marker comprises an SNP locus 1 and an SNP locus 2; the SNPlocus 1 is located at the base 30654770 of a rice chromosome 11, and the molecular marker polymorphism is A / T; the SNP locus 2 is located at the base 30655545 of the rice chromosome 11, and the molecular marker polymorphism is C / G. The molecular marker is high in Pi1 gene site specificity, an application method of the SNP molecular marker is convenient to implement, the detection result is accurate and reliable, the marker can be used for identification and assisted breeding of the rice Pi1 gene, and the problems that traditional breeding is low in efficiency and long in cycle can be solved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a KASP molecular marker for detecting rice blast resistance broad-spectrum gene Pil and its application. Background technique [0002] Rice (Oryza sativa L.) is an important food crop. Rice blast is a disease caused by ascomycetes. Since the 1990s, the average annual rice blast area in my country has exceeded 40 million hm2, and the loss has reached 100,000 kg. Therefore, rice blast is a serious obstacle to rice production in my country. , is also a difficult problem in the process of rice breeding. Decades of practical experience in production have shown that identification of blast-resistant genotypes of main rice varieties and breeding of resistant varieties are the most economical and effective means to solve rice blast. [0003] As of March 2015, 84 major genes have been mapped in 69 rice blast resistance loci, and 24 of them have been cloned. These genes are locat...

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 郭嗣斌韦宇李孝琼陈颖刘开强
Owner 广西马坪壮欢科技有限公司
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