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SNP molecular marker in close linkage with bacterial blight resistance gene Xa7 and applications of SNP molecular marker

A bacterial blight resistance and molecular marker technology, which can be used in recombinant DNA technology, microbial assay/test, DNA/RNA fragments, etc. Design gene function markers and other issues to achieve the effect of facilitating high-throughput rapid detection, eliminating aerosol pollution, and accurate detection

Active Publication Date: 2017-05-10
HUAZHI RICE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, since the Xa7 gene has not been cloned, gene function markers could not be designed
As a result, the bacterial blight resistance gene Xa7 has not been well utilized in traditional rice breeding

Method used

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  • SNP molecular marker in close linkage with bacterial blight resistance gene Xa7 and applications of SNP molecular marker
  • SNP molecular marker in close linkage with bacterial blight resistance gene Xa7 and applications of SNP molecular marker
  • SNP molecular marker in close linkage with bacterial blight resistance gene Xa7 and applications of SNP molecular marker

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Experimental program
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Embodiment 1

[0034] This example is used to illustrate the application of the molecular marker provided by the present invention in the detection of bacterial blight resistance gene Xa7, and the specific steps are as follows:

[0035] 1. According to the molecular markers described in the present invention, design primer combinations that can be used for high-throughput detection of Xa7 resistance gene in rice materials by using KASP reaction, as shown in Table 1.

[0036] 2. Genomic DNA was extracted from rice leaves by simplified CTAB method.

[0037] ①Put the sample into a 2.0mL Tube, add two steel balls and 750μL CTAB solution in advance, shake the homogenized sample for 1.5min;

[0038] ②Concussion heating at 65℃ for 0.5-1h;

[0039] ③ Cool to room temperature, add 750mL chloroform:isoamyl alcohol (24:1) solution in the fume hood and mix well;

[0040] ④ Centrifuge at 12000rmp for 10min, take about 500mL supernatant and transfer it to a new 1.5mL centrifuge tube;

[0041] ⑤ Add an ...

Embodiment 2

[0056] 1. Genetic position verification of molecular markers

[0057] Nine genome-wide markers with polymorphisms in the parents and two markers linked to Xa7 in the present invention were used to verify the genetic position. The two molecular markers linked to Xa7 in the present invention were both mapped to chromosome 6 82.7 cM position, such as image 3 shown.

[0058] 2. Phenotypic verification of molecular markers

[0059] The genetic phenotype of the two Xa7-linked markers in the present invention was verified by using 120 F2 hybrid populations of the donor parent Hua 1228S and the recurrent parent R608. At the rice booting stage, 120 individual plants of the F2 population and 2 parental varieties were inoculated with the Guangdong bacterial blight strain GD1358, and the disease was investigated 21 days later. The consistency between the phenotype data and the genotype was more than 90%, which verified the method of the present invention again. Feasibility and accurac...

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Abstract

The invention relates to molecular markers, and in particular discloses an SNP molecular marker K_060569 in close linkage with a bacterial blight resistance gene, wherein the sequence of the SNP molecular marker K_060569 is shown as SEQ ID No.1, and the basic group at the 61st bp site is C or G. The invention further provides a primer combination for carrying out Xa7 resistance gene detection on a rice material at high throughput by adopting the KASP reaction, and applications of the molecular marker and the primer combination in detecting the bacterial blight resistance gene Xa7 and in disease resistance assistant breeding of rice. According to the scheme, the KASP technology is adopted for carrying out genetic typing on the searched SNP molecular marker, the Xa7 gene can be rapidly and accurately detected, and the gene transferring breeding efficiency can be greatly improved; during the detection process, the operations such as digestion, electrophoresis and sequencing are not needed, so that the operation is simple and convenient, the high-throughput rapid detection is facilitated, the aerosol pollution of the PCR product, the pollution of EB to the environment, and the harm of formaldehyde to the human body are completely eradicated.

Description

technical field [0001] The invention relates to a molecular marker, in particular to a SNP molecular marker closely linked with a bacterial blight resistance gene. Background technique [0002] Rice bacterial blight has physiological race specialization, and the resistance of varieties is basically controlled by the main resistance gene in the nuclear genome. The hybrid progenies analyzed their resistance to Japanese flora, and after a dominant resistance gene was identified and named, the research on the identification and excavation of rice bacterial blight resistance genes has not stopped. After 1975, the International Rice Research Institute (using the Philippine physiological race) successively identified four disease resistance genes (Zhang Qi, 2007). Researchers in other countries also identified a number of resistance genes one after another, but due to the strains used by scientists from various countries different, the lack of comparability of its identification r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 郑秀婷彭佩江南李为国熊艳文李继明
Owner HUAZHI RICE BIO TECH CO LTD
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