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CAPS molecular marker and method for identifying rice varieties and application of molecular marker

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: 2019-05-10
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 and method for identifying rice varieties and application of molecular marker

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Effect test

Embodiment 1

[0033] Example 1 Identification of rice CAPS molecular marker MAIF1-SNP1 and detection primers

[0034] The inventors found in previous studies that there is a molecular marker MAIF1-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 394bp, and there is a T←→C SNP site difference, it is speculated that the above molecular markers can be used to identify rice indica and japonica varieties. The analysis found that the japonica rice has a restriction endonuclease BsrF I restriction endonuclease recognition site in the adjacent sequence of the SNP site: GCCGGC, while the SNP mutation GTCGGY at the restriction endonuclease recognition site in indica rice cannot be recognized by the restriction endonuclease BsrF I enzyme digestion recognition, so the CAPS molecular marker method can be used to detect and identify rice indica and japonica rice varieties. When MAIF1-SNP1 is marked with BsrF ...

Embodiment 2

[0040] Embodiment 2 identifies the method for rice indica and japonica varieties

[0041] The experimental materials used in the method for identifying rice indica and japonica varieties of the present invention include common wild rice and Nivara wild rice; the indica varieties are respectively 93-11, Bianjiao Nante, Guangluai No. 4, Zhenshan 97, Minghui 63 , Huangsizhan, Huanghuazhan; japonica rice varieties were Nipponbare, Taichung 65, Zhonghua 11, Nongken 58, Yunjing 23, Ewan 3, and Jindao 1. Specific steps are as follows:

[0042] 1. Genomic DNA extraction from rice samples

[0043] (1) Take about 0.5g 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 2mL centrifuge tube, add 800μL 2×CTAB extraction buffer solution, mix well, and take a water bath at 65°C for 30 minutes;

[0044] (2) Add an equal volume of chloroform, isoamyl alcohol and absolute ethanol (76:4:20), shake for 10 m...

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Abstract

The invention discloses a CAPS molecular marker and method for identifying rice varieties and application of the molecular marker. The nucleotide sequence of the CAPS molecular marker in oryza sativasubsp japanica is shown as SEQ ID NO:1, and the sequence of the CAPS molecular marker in oryzasativa subsp indica is shown as SEQ ID NO:2. The invention also provides a primer group for detecting theCAPS molecular marker, and the sequence of the primer group is shown as SEQ ID NO:3-SEQ ID NO:4. The provided CAPS molecular marker has high specificity and can be used for conveniently and accuratelydistinguishing germplasm resources of oryzasativa subsp indica from those of oryza sativa subsp japanica. The provided CAPS molecular marker only requires PCR-combined enzyme digestion in actual application and has the advantages of being 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 CAPS molecular marker and method for identifying rice varieties and applications thereof. Background technique [0002] Rice varieties are mainly cultivated in Asia, and are divided into indica rice (Oryza sativa subsp Indica) and japonica rice (Oryza sativa subsp Japanica) according to a series of phenotypic differences. my country has developed and cultivated thousands of indica rice according to different regional planting conditions and production needs. and japonica rice varieties. 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 p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 张群宇袁健铭王曼周峰
Owner SOUTH CHINA AGRI UNIV