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CAPS molecular marker method for identifying rice varieties and application

A molecular marker and rice technology, applied in the field of plant biology, can solve problems such as the great influence of experience, and achieve the effect of high throughput, high specificity and low cost

Active Publication Date: 2018-01-26
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The identified traits mainly include three aspects, namely, plant traits, ear traits and grain traits, but the actual production and application are greatly influenced by the experience of the appraiser

Method used

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  • CAPS molecular marker method for identifying rice varieties and application

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Effect test

Embodiment 1

[0030] Example 1 Molecular markers and detection primers for identifying rice

[0031] The inventors found in previous studies that there is a molecular marker V14-SNP1 in rice, its sequence in japonica rice is shown in SEQ ID NO: 1, and the sequence in indica rice is shown in SEQ ID NO: 2, both of which are 440bp, and at 222bp, there is a SNP site difference of G←→C. It is speculated that the above molecular markers can be used to identify rice indica and japonica varieties. The analysis found that because the japonica rice has a restriction endonuclease in the adjacent sequence of the SNP site Hha I restriction endonuclease recognition site: GCGC, and the SNP mutation GCGG of indica rice at this restriction endonuclease Hha I restriction restriction recognition, therefore can detect and identify rice indica rice and japonica rice varieties by CAPS molecular marker method, when using Hha After the restriction enzyme recognition molecular marker V14-SNP1, the japonica ...

Embodiment 2

[0037] Example 2 CAPS molecular marker method for identifying rice indica and japonica varieties

[0038]The experimental materials for the CAPS molecular marker method used to identify rice indica and japonica varieties in the present invention include common wild rice and Nivara wild rice; the indica varieties are 93-11, Banjiao Nante, Guangluai 4, Zhenshan 97, respectively. Minghui 63, Huangsizhan, and Huanghuazhan; the japonica rice varieties were Nipponbare, Taichung 65, Zhonghua 11, Nongken 58, Yunjing 23, Ewan 3, and Jindao 1. Specific steps are as follows:

[0039] 1. Genomic DNA extraction from rice samples

[0040] (1) Take about 0.5 g of rice leaves, cut them into pieces, put them into a pre-cooled mortar, add liquid nitrogen, grind the leaves into powder, put them into a 2 mL centrifuge tube, add 800 μL 2×CTAB to pump Extract the buffer, mix well, and bathe in 65°C water for 30 minutes;

[0041] (2) Add an equal volume of chloroform, isoamyl alcohol and absolute...

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Abstract

The invention discloses a CAPS molecular marker method for identifying rice indica and japonica varieties and application. The method comprises the following steps: S1. extracting rice sample genomicDNA; S2. carrying out PCR amplification on the genomic DNA in step S1 with a primer of claim 3; S3. subjecting a PCR product in step S2 to electrophoresis after (i) Hha I digestion to obtain a CAPS marker; and S4. judging an electrophoresis result: if a 440bp electrophoresis band is obtained, identifying the rice sample as indica, and if a 220bp electrophoresis band is obtained, identifying the rice sample as japonica. The CAPS molecular marker method disclosed by the invention has high specificity, and can conveniently and accurately distinguish rice germplasm resources of indica and japonicafor rice cultivation varieties. Meanwhile, the molecular marker provided by the invention only needs PCR combined with digestion in a practical application, and is low in cost, high in flux and highin specificity, thereby being very applicable to production practice.

Description

technical field [0001] The invention belongs to the field of plant biotechnology. More specifically, it relates to a method and application of a CAPS molecular marker for identifying rice varieties. Background technique [0002] Rice cultivars are mainly cultivated in Asia, and are divided into indica ( Oryza saliva subsp Indica ) and Japonica ( Oryza saliva subsp Japan ), my country has developed and cultivated thousands of indica and japonica rice varieties according to different regional planting conditions and production needs. To ensure high and stable yields, the seed purity of rice varieties is one of the most critical factors. Traditionally, the identification of the purity of rice varieties is carried out at different growth stages such as seedling stage, heading stage, and ripening stage. The identified traits mainly include three aspects, namely, plant traits, panicle traits and grain traits, but the actual production and application are greatly influenced...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 张群宇刘耀光周峰王曼薛德星裴蕾江燕翔赵秀彩祝钦泷
Owner SOUTH CHINA AGRI UNIV