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Microsatellite molecular marker and method for authenticating rice varieties and application of mcriosatellite molecular marker and method

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

Active Publication Date: 2019-04-16
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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  • Microsatellite molecular marker and method for authenticating rice varieties and application of mcriosatellite molecular marker and method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Identification of rice microsatellite molecular marker WRKY10 and detection primers

[0034] The inventor’s previous research found that there is a molecular marker WRKY10 in rice, and its sequence in japonica rice is shown in SEQ ID NO: 1, and in indica rice is shown in SEQ ID NO: 2, and exists at 83 bp. The difference in the number of tandem repeats of a CGG microsatellite is speculated that the above-mentioned molecular markers can be used to identify rice indica and japonica varieties. The analysis found that since japonica rice has 6 CGG repeats at this locus, while indica rice has only 2 CGG repeats at this locus, microsatellite molecular marker methods can be used to detect and identify rice indica and japonica rice varieties. The above DNA fragments were amplified by PCR. The japonica rice fragment was a 129bp DNA fragment, while the indica rice fragment was a 117bp DNA fragment, which was used to identify the indica and japonica rice varieties.

[0035] A...

Embodiment 2

[0040] Example 2 Method for identifying indica and japonica rice varieties

[0041] The experimental materials used in the method for identifying indica and japonica rice varieties of the present invention include common wild rice and Nivara wild rice; the indica varieties are 93-11, Aijiao Nante, Guangluai 4, Zhenshan 97, Minghui 63 , Huang Si Zhan, Huang Hua Zhan; Japonica rice varieties are Nipponbare, Taichung 65, Zhonghua 11, Nongken 58, Yunjing 23, Ewan 3, Jindao 1. Specific steps are as follows:

[0042] 1. Extraction of genomic DNA from rice samples

[0043] (1) Take about 0.5g rice leaves, cut them into small pieces, put them in a pre-cooled mortar, add liquid nitrogen, grind the leaves into powder, put them in a 2mL centrifuge tube, add 800μL 2×CTAB extraction buffer Liquid, mix well, water bath at 65℃ for 30min;

[0044] (2) Add equal volume of chloroform, isoamyl alcohol and absolute ethanol (76:4:20), shake for 10 minutes, and mix well;

[0045] (3) Centrifuge for 12 min...

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Abstract

The invention discloses a microsatellite molecular marker and a method for authenticating rice varieties and application of the mcriosatellite molecular marker and the method. The nucleotide sequenceof the microsatellite molecular marker in japonica rice is shown in SEQ ID NO:1, and the sequence of the microsatellite molecular marker in indica type rice is shown in SEQ ID NO:2. The invention further provides a primer group used for detecting the microsatellite molecular marker. The sequences of the primer group are shown in SEQ ID NO:3-SEQ ID NO:4. The microsatellite molecular marker is highin specificity and can accurately and conveniently conduct germplasm resource distinguishing of indica type rice and japonica rice on the rice cultivated varieties. In actual application of the microsatellite molecular marker, only PCR and enzyme digestion are combined, and the microsatellite molecular marker is low in cost, high in flux and specificity and suitable for production practice.

Description

Technical field [0001] The invention relates to the field of plant biotechnology, in particular to a microsatellite molecular marker for identifying rice varieties, a method and application thereof. Background technique [0002] Rice varieties are mainly Asian cultivated rice, and are divided into indica rice based on a series of phenotypic differences ( Oryza sativa subsp Indica ) And japonica rice ( Oryza sativa subsp Japanica ), my country has developed and cultivated thousands of indica and japonica varieties according to different geographical planting conditions and production needs. To ensure high and stable yields, seed purity of rice varieties is one of the most critical factors. The purity of rice varieties is traditionally identified in different growth periods such as seedling stage, heading stage, and ripening stage. There are three main aspects of the identified traits, namely plant traits, ear traits and grain traits, but the actual production and application...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 赵秀彩丁智张群宇袁健铭
Owner SOUTH CHINA AGRI UNIV