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SNP molecular marker for identifying aquilaria sinensis, aquilaria yunnanensis and aquilaria malaccensis and application of marker

A technology of molecular markers and agarwood, which is applied in the field of molecular biology detection of wood species identification, can solve the problems of subjective identification deviation and morphological differences, and achieve the effect of simple operation and objective identification process

Pending Publication Date: 2019-03-19
HANGZHOU KMB BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Morphological identification is a commonly used method in the identification of agarwood, but the shape characteristics are easily affected by habitat, climate, physiological conditions, etc., which often lead to deviations in subjective identification, and it is difficult to accurately identify only by morphological differences

Method used

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  • SNP molecular marker for identifying aquilaria sinensis, aquilaria yunnanensis and aquilaria malaccensis and application of marker
  • SNP molecular marker for identifying aquilaria sinensis, aquilaria yunnanensis and aquilaria malaccensis and application of marker
  • SNP molecular marker for identifying aquilaria sinensis, aquilaria yunnanensis and aquilaria malaccensis and application of marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Acquisition of SNP Molecular Markers Used to Identify Agarwood, Yunnan Agarwood, and Malayan Agarwood

[0054] The 5.8, 18 and 28S rRNA genes on rDNA are highly conserved, that is, there are extensive heterologous homology. And because the ITS region does not add mature ribosomes, the natural selection pressure on the ITS fragment during the evolution process is very small, so it can tolerate more mutations. In the vast majority of eukaryotes, sequence polymorphisms are very extensive, and even two species that are very closely related can show differences in ITS sequences, showing recent evolutionary characteristics. This characteristic makes ITS suitable for the molecular identification of species and the analysis of phylogenetic relationship with obvious differences between species or within a genus. Because the sequence analysis of ITS can substantially reflect the base pair differences between genera and species, and the sequence fragments of ITS are sma...

Embodiment 2

[0067] Example 2 Application of SNP Molecular Markers of Agarwood in the Identification of Agarwood, Yunnan Agarwood, and Malay Agarwood by HRM-PCR Amplification Method

[0068] In this example, on the basis of obtaining the SNP sites of Agarwood, Yunnan Agarwood, and Malayan Agarwood, specific upstream primers F2, downstream primers R2 and Probes and primers were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. Using the specific primers, the HRM-PCR amplification detection was carried out on the samples of Agarwood terrescens, samples of Agarwood malayi and samples of Agarwood yunnanensis. Specifically, the upstream primer F, downstream primer R and probe are respectively:

[0069] Upstream primer F2: 5'-AAGGAACTTGATCACATAATATGTT-3' (SEQ ID NO.6)

[0070] Downstream primer R2: 5'-TCATTTTAGATTCGTTTCAACGC-3' (SEQ ID NO.7)

[0071] Using the genomic DNA of 3 Aquilaria species as templates, the 20 μL PCR reaction system includes:

[0072]

[0073] Fluorescence PCR ...

Embodiment 3

[0081] Example 3 Application of Agarwood SNP Molecular Markers in the Identification of Agarwood Agarwood, Agarwood Yunnan, and Agarwood Malayi by ARMS-PCR

[0082] In this example, on the basis of obtaining the SNP sites of Agarwood tulsifolia, Agarwood yunnanensis and Agarwood malayi, specific upstream primers and downstream primers were designed using PrimerPremier 5.0 for the gene sequences containing the SNP sites. The primers were composed of Synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. Utilize the specific primers and probes to carry out routine PCR amplification detection on the samples of Agarwood edulis, samples of Agarwood malayi and samples of Agarwood yunnanensis. Specifically, the upstream primer F, the downstream primer R and are respectively:

[0083] Upstream primer FP: 5'-CTTGTGGCCGTCATAACCAAACC-3' (SEQ ID NO.8)

[0084] Agarwood downstream primer TR: 5'-CGTTTCAACGCTTGATCTCA-3' (SEQ ID NO.9)

[0085] Downstream primer YR of Agarwood yunnanensis: 5...

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Abstract

The invention discloses an SNP molecular marker for identifying aquilaria sinensis, aquilaria yunnanensis and aquilaria malaccensis and an application of the marker. A system for rapidly identifying the aquilaria sinensis, the aquilaria yunnanensis and the aquilaria malaccensis is established based on the SNP molecular marker, and thus the propose of rapidly and accurately detecting the aquilariasinensis, the aquilaria yunnanensis and the aquilaria malaccensis is realized, so that the marker is of great significance for species inspection at ports and the like.

Description

technical field [0001] The invention relates to the field of molecular biology detection for identification of wood species, in particular to SNP molecular markers for identifying agarwood, agarwood yunnanensis and agarwood malayi and applications thereof. Background technique [0002] Agarwood, also known as "Shen Shuixiang", "Shui Chenxiang", is written as "Agarwood" (Shen, same as Shen) in the old saying. The "Shen" in "Shen Tanlong Musk" often said in ancient times refers to agarwood. Agarwood incense is elegant and rare, and has been listed as the top of all incense since ancient times. [0003] Unlike sandalwood, agarwood is not a kind of wood, but a solid condensate that is "knotted" by a special type of fragrant tree, which is a mixture of oil (resin) components and woody components. The wood itself of this kind of incense tree has no special fragrance, and the wood is relatively soft. According to current research, several trees of the genus Agarwood in the famil...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2600/166C12Q2531/113C12Q2527/107C12Q2563/107
Inventor 林源吉丁佳女郑宜文
Owner HANGZHOU KMB BIOTECH
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