SNP molecular marker used for identifying camellia nitidissima and application thereof

A molecular marker, golden camellia technology, applied in the field of molecular biology, can solve the problems of complicated technology and high cost

Inactive Publication Date: 2015-01-21
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current SNP detection and analysis methods, such as: DNA direct sequencing, gene chip technology, array hybridization analysis, high-performance denaturing liquid chromatography detection, etc. (Jehan and Lakhanpaul, 2006), but due to cumbersome technology and high cost, its application is limited

Method used

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  • SNP molecular marker used for identifying camellia nitidissima and application thereof
  • SNP molecular marker used for identifying camellia nitidissima and application thereof
  • SNP molecular marker used for identifying camellia nitidissima and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] The acquisition of embodiment 1 golden camellia SNP molecular marker

[0021] Granule-bound starch synthase I (GBSSI) is a key enzyme for the synthesis of amylose in endosperm and other plant storage organs. Its function is conserved and it is a single copy in most plant families. Studies have shown that the GBSSI gene in Camellia is a single copy, and has a high sequence variation rate (the variation between different species is about 5%), which is very useful for studying the phylogeny of groups such as Camellia with many closely related species. Relationships have significant value (Yang et al., 2006).

[0022] 1. According to the primers and amplification conditions of the GBSSI gene published by Yang Junbo et al. (2006), the genomic DNA of 17 Camellia species in Table 1 was used as a template for PCR amplification, and the amplified product was passed through 1.5% agar Glycogel electrophoresis detection and sequencing after qualified observation by UV gel imaging ...

Embodiment 2

[0031] Example 2 Application of Camellia SNP Molecular Markers

[0032] This example establishes the dCAPS molecular marker system based on the SNP sites of Camellia japonica, so as to quickly identify the rare and endangered species in the world-Camellia japonica.

[0033] It includes the following steps: 1) extracting the genomic DNA of the plant to be tested; 2) using the genomic DNA of the plant to be tested as a template, using the dCAPS primer (5'-CTGTGGACGCAAACATCCACTTGAT-3') and the corresponding downstream primer (5'-CCATGTATTTCTTGCCAGTGCCCT-3 '), PCR amplification of Camellia japonica GBSSI gene; 3) restriction endonuclease BstXI was used to digest the PCR amplification product (the specific method was the same as in Example 1), and the digested product was subjected to agarose gel electrophoresis, and the electrophoresis results showed 2 strips band, the plant to be tested is identified as Camellia japonica species.

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Abstract

The invention provides an SNP molecular marker used for identifying camellia nitidissima and an application thereof. The SNP is located at a 141st-position basic group of a GBSSI gene, represented as SEQ ID No.1, of the camellia nitidissima. If the basic group is T, a plant is identified as the camellia nitidissima. In the invention, a dCAPS molecular marker system is firstly established on the basis of the SNP locus of the camellia nitidissima, and analysis is carried out to a gene sequence of the GBSSI gene of the camellia nitidissima through a bioinformatic method for screening the SNP locus. A proper mispairing primer is designed for PCR amplification. A proper restriction endonuclease is selected for enzyme digestion. Finally the SNP is converted into the dCAPS molecular marker for detection. A quick identification method suitable for port check of the camellia nitidissima is established.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a SNP molecular marker for identifying camellia japonica and its application. Background technique [0002] Camellia nitidissima Chi (Camellia nitidissima Chi) is an evergreen shrub belonging to the Camellia group (Sect.chrysantha) of the family Theaceae. It has important horticultural value because of its yellow corolla, and is known as the "Queen of Camellia . In recent years, its natural population has declined sharply due to habitat loss, overharvesting of endangered species resources, and illegal export trade. Camellia camellia is listed as a first-class national key protected plant and one of the most endangered plant species in the IUCN Red List. Therefore, establishing an accurate identification method for Camellia japonica plants is the key to the conservation and research of its biodiversity. [0003] Single nucleotide polymorphism (single nucleotide polymorphism, SN...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 宋云赵竹徐晗
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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