Primer for identifying rhizoma atractylodis medicinal species by using chloroplast genes, and applications thereof

A technology of chloroplast and Atractylodes genus, which is applied in the field of using chloroplast genes to identify primers for medicinal species of Atractylodes genus, which can solve the problems of unsatisfactory performance, unclear species boundaries, and low resolution

Active Publication Date: 2020-04-14
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Abstract
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Problems solved by technology

However, the performance of these markers at the species and subspecies levels is unsatisfactory due to unclear species boundaries and low resolution

Method used

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  • Primer for identifying rhizoma atractylodis medicinal species by using chloroplast genes, and applications thereof
  • Primer for identifying rhizoma atractylodis medicinal species by using chloroplast genes, and applications thereof
  • Primer for identifying rhizoma atractylodis medicinal species by using chloroplast genes, and applications thereof

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Embodiment 1

[0044] 1. The genomes of Atractylodes Rhizoma Atractylodes Rhizome, Atractylodes Rhizoma Atractylodes Rhizome and Atractylodes Rhizoma Atractylodes were extracted, sequenced by NGS technology, and the chloroplast genome sequences of Atractylodes Rhizome, Atractylodes Rhizome and Atractylodes Rhizome were obtained, and data analysis was performed on the sequences by software.

[0045] Data difference analysis method:

[0046] The present invention uses REPuter software to identify four types of repeat sequences, including forward repeats, palindromic repeats, inverted repeats and complementary repeats; the minimum repeat unit size is set to 30bp, and the critical value of similarity between repeat units is set to 90%.

[0047] Use MISA software to predict simple repeats (SSR), and the thresholds are as follows: ① When the number of repeating unit nucleotide residues is 1, the number of repeating units is at least 8; ② The number of repeating unit nucleotide residues is 2-3 3, ...

Embodiment 2

[0093] In order to further verify the reliability of the above molecular markers, Atractylodes Rhizoma Atractylodes Rhizome and Atractylodes Rhizoma Atractylodis selected plant materials from 3 origins, and 6 individuals (Table 10) were collected from each origin. Atractylodes macrophylla selected plant materials from 2 origins, and collected 2 individuals (Table 10) from each origin. Total DNA extraction was performed on all 40 individuals. PCR amplification was performed with the primer pair cz11_F / cz11_R. The corresponding PCR products were sequenced by Sanger sequencing. The alignment results of the 40 PCR product sequences are shown in ( Figure 5 )middle. The 36 individuals of Atractylodes Rhizoma Atractylodes Rhizome and Atractylodes Rhizoma Atractylodes Rhizome all showed expected marker sequences (Table 10) and were successfully verified. The 4 individuals of Atractylodes macrophylla also showed the expected marker sequence (Table 10) and were successfully verifie...

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Abstract

The invention discloses a primer for identifying rhizoma atractylodis medicinal species of Atractylodes macrocephala, Atractylodes lancea and Atractylodes chinensis by using chloroplast genes, and applications thereof. According to the invention, a sequence represented by SEQ ID NO.1 / SEQ ID NO.2 is used as a primer to amplify the chloroplast genome sequences of Atractylodes macrocephala, Atractylodes lancea and Atractylodes chinensis, and the comparison results after PCR strip sequencing show that the target strip of the Atractylodes lancea has 5 bps tatat insertion mutation, and the target strip of the Atractylodes macrocephala has 6 bps tcttac insertion mutation; and the chloroplast genome molecular marker successfully distinguishes three species of Atractylodes macrocephala, Atractylodes lancea and Atractylodes chinensis, and provides a technical means for genetic diversity, phylogenetic evolution and taxonomy research of rhizoma atractylodis plants.

Description

technical field [0001] The method relates to the field of plant molecular biology, in particular to a primer for identifying medicinal species of the genus Atractylodis by using chloroplast genes and its application. Background technique [0002] There are 7 species of Atractylodes (Asteraceae) distributed in China, Korea and Japan. The medicinal properties of many Atractylodes species have long been known. According to clinical use, its dried rhizomes can be used as two kinds of Chinese herbal medicines, namely Atractylodes Rhizoma Atractylodes Rhizome and Atractylodes Rhizoma Atractylodes Rhizome. In "Chinese Pharmacopoeia", "Korean Pharmacopoeia" and "Japanese Pharmacopoeia", the dried rhizome of Atractylodes atractylodes is called Atractylodes atractylodes, and the dried rhizomes of Atractylodes atractylodes and Atractylodes atractylodes are called Atractylodes atractylodes. [0003] The base plants of traditional Chinese medicine Atractylodes atractylodes can be divid...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6869C12N15/11
CPCC12Q1/6895C12Q1/6869C12Q2600/13C12Q2531/113C12Q2535/101Y02A50/30
Inventor 刘昶张慧王立强姜梅
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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