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est-ssr marker identification method and primers of traditional Chinese medicine Xihuangcao species

A technology of Xihuangcao and traditional Chinese medicine, which is applied in the field of molecular markers and medicinal material identification, can solve the problem of lack of information in the database, and achieve the effect of facilitating dry sample analysis, simple method, and high timeliness

Inactive Publication Date: 2018-05-04
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research on Xihuangcao rarely involves the molecular field, and the information in various databases is very scarce

Method used

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  • est-ssr marker identification method and primers of traditional Chinese medicine Xihuangcao species
  • est-ssr marker identification method and primers of traditional Chinese medicine Xihuangcao species
  • est-ssr marker identification method and primers of traditional Chinese medicine Xihuangcao species

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 The development of the EST-SSR core primer set based on the transcriptome sequence of Xihuangcao

[0034] 1. Construct the transcriptome information database of Xihuangcao and Xianwen Xiangchacai

[0035] The same amount of roots, stems, leaves, and flowers of Xihuangcao and Xianwen Xiangchacai were collected in the medicine garden of Guangdong Provincial Institute of Traditional Chinese Medicine. After mixing, RNA was extracted and sequenced. After obtaining massive data, use the GO classification (Gene Ontology) method to compare all spliced ​​unigenes with the NCBI non-redundant protein database, remove redundant sequences, and set the E-value to 10 -6 . The annotation information of uinigenes of all non-redundant sequences was then extracted using Blast2GO software. Finally, all unigenes with non-redundant sequences were functionally classified (by molecular function, cellular components, biological process, metabolic process, etc.) with the online so...

Embodiment 2

[0063] Example 2 Utilize EST-SSR markers to identify the species of Chinese medicine Xihuangcao

[0064] 1. DNA extraction

[0065] Use the improved CTAB method to extract the DNA from the leaves of the above-mentioned 8 genuine and counterfeit plants of Xihuangcao, quantify them with a UV spectrophotometer, dilute to 50ng / μl, and store them at 4°C or -20°C until use;

[0066] Concrete steps are with embodiment 1.

[0067] 2. PCR amplification

[0068] Using the DNA extracted in step 1 as a template, the 24 pairs of EST-SSR core primers developed in Example 1 were used for PCR amplification. The amplification method was the same as in Example 1, and amplified products with different lengths and different fluorescences were obtained.

[0069] 3. Detection and analysis

[0070] PCR products of different lengths and different fluorescences were mixed in equal proportions, capillary electrophoresis and typing were carried out with a 3730xl DNA sequencer (ABI, USA), and then the...

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Abstract

The present invention relates to a method utilizing an EST-SSR marker for identification of traditional Chinese medicine serrate rabdosia herb varieties , the method comprises the following steps: (1) extracting genomic DNA of a sample to be tested; (2) using the DNA extracted by the step (1) as a template, and using a serrate rabdosia herb transcriptome sequence-based EST-SSR core primer group for PCR amplification; and (3) typing amplified products on the basis of sequencing; the EST-SSR core primer group comprises at least 10 pairs of primers of the 24 pairs of primers, each pair of the primers comprise an upstream primer and a downstream primer, and the nucleotide sequences are SEQ ID NO. 1-SEQ ID NO.48. The method can achieve effective and rapid identification of common serrate rabdosia herb medicine-based source varieties and adulterants, and is accurate and reliable in results, simple, easy, high in timeliness, and suitable for the serrate rabdosia herb medicinal material variety identification and variety protection in the market and the production.

Description

technical field [0001] The invention relates to the technical field of molecular markers and identification of medicinal materials, in particular to a method and primers for using EST-SSR markers to identify varieties of Chinese medicine Xihuangcao. Background technique [0002] Xihuangcao, a traditional Chinese medicine, is a herbal medicine commonly used by the people. It is a plant of the genus Isodon in the family Lamiaceae. It is mainly produced in Hunan, Hubei, Sichuan, Yunnan, Jiangxi, Guangdong, Guangxi, Fujian and other provinces south of the Yangtze River. . Xi was born by the damp stream in the valley, and the fresh leaves are mashed with yellow juice, so it gets its name. Its nature and flavor are bitter, sweet, and cold, and it returns to the liver and gallbladder meridian. Xihuangcao is widely used clinically in Guangdong, mainly for the treatment of acute jaundice hepatitis, acute cholecystitis, damp-heat dysentery, enteritis, bruises and bruises, and health...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/158
Inventor 黄珊珊王瑛曾少华胡伟明黄鹤莫小路曾庆钱袁亮
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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