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Method for identifying traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof on the basis of ITS2 sequence gene fragments

A gene fragment and plant technology, applied in the field of identification of the original plant species of traditional Chinese medicinal materials, can solve the problems affecting the clinical efficacy and drug safety of raspberries, differences in chemical composition and pharmacological effects, etc., and achieves easy control, high accuracy, and applicability. wide effect

Inactive Publication Date: 2018-01-05
LISHUI AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the traits of raspberry and Rubus are similar, there are obvious differences between raspberry and these phytochemical components and pharmacological effects, and the mixed use has seriously affected the clinical efficacy and drug safety of raspberry. It is particularly important to accurately identify
[0004] Due to the limitations of traditional identification methods in the identification of Chinese medicinal materials, in order to ensure the safety and effectiveness of clinical use of Chinese medicinal materials, it is necessary to use new technologies and methods to supplement them.

Method used

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  • Method for identifying traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof on the basis of ITS2 sequence gene fragments
  • Method for identifying traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof on the basis of ITS2 sequence gene fragments
  • Method for identifying traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof on the basis of ITS2 sequence gene fragments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Distinguishing the original object of Twisted chinensis (Raspberry palmatum) and the leaves of the same genus and similar easily mixed species of Sorghum japonica, raspberry, pomegranate, Mao berry and Hanberry

[0033] 1 provide samples

[0034] A total of 67 leaves samples of the original root of Twister chinensis (Raspberry palmatum) and its similar genus and hybrid species Sorghum japonica, Raspberry, Pompon, Maoberry and Hanberry were collected from different places of origin (see Table 1 for details)

[0035] Table 1. Leaf samples of the original stock of the common twisted root (Raspberry palmatum) and its genus and similar hybrid species Sorghum, Raspberry, Pompon, Maoberry and Hanberry

[0036]

[0037] 2 Genomic DNA Extraction

[0038] Dry the leaf samples in Table 1 with color-changing silica gel, take about 20 mg of leaves, grind for 2 min (30 times / second) with a grinder (Retsch MM400, Germany), and then extract the total DNA with a plant DNA ...

Embodiment 2

[0059] Example 2 Distinguish the roots of the original object of the genus Twisted Root (Raspberry palmatum) and the roots of the same genus and similar easily mixed species Sorghum japonica, raspberry and pomegranate

[0060] 1 provide samples

[0061] Collected 8 roots of the original species (Raspberry palmatum) and its similar genus, Sorghum, Raspberry, and Pamponga from different origins (see Table 3 for details)

[0062] Table 3 The root sample information table of the original species (Raspberry palmatum) and its similar genus Sorghum, Raspberry and Pontus chinensis

[0063]

[0064] 2 Genomic DNA Extraction

[0065] The root samples in Table 3 were dried with color-changing silica gel, and about 50 mg of the roots were taken, and the total DNA was extracted with a plant DNA extraction kit (Tiangen Biotech Co., China) after grinding for 2 min (30 times / second) with a grinder (Retsch MM400, Germany). Water bath at 65°C for 1 h, and the remaining steps were carried o...

Embodiment 3

[0087] Example 3 Distinguish the original object of Twisted chinensis (Raspberry palmatum) and the fruits of the same genus and similar easily mixed species of sorghum, raspberry, pomegranate and white leaf berry

[0088] 1 provide samples

[0089] A total of 9 copies of the original species (Raspberry palmatum) and the similar and easily mixed species Sorghum japonica, Raspberry, Pompon and White leaf berry of the same genus were collected from different origins (see Table 5 for details)

[0090] Table 5 The fruit sample information table of the original species (Raspberry palmatum) and its similar genus, Sorghum, Raspberry, Pompon and White-leaf Berry

[0091]

[0092] 2 Genomic DNA Extraction

[0093] Dry the root samples in Table 3 with color-changing silica gel, take about 80 mg of fruit samples, grind for 2 min (30 times / second) with a grinder (Retsch MM400, Germany), and then extract the total DNA with a plant DNA extraction kit (Tiangen Biotech Co., China) , in a ...

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Abstract

The invention relates to the field of traditional Chinese medicinal material original plant species identification technology, specifically to a method for identifying leaf, root and fruit of traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof by the utilization of ITS2 fragments of ribosome DNA and application thereof. The method for identifying traditional she medicine Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof on the basis of ITS2 sequence gene fragment comprises the following steps: 1) extraction of multiple rubus plant leaf, root and fruit DNA samples; 2) PCR amplification of fragments containing ITS2 sequence; 3) assembling of PCR amplification fragments; 4) comparison of ITS2 sequences of multiple rubus species; and 5) construction of a phylogenetic tree on the basis of ITS2 sequence. The method of the invention has extensive applicability, is simple and convenient to operate,is easy to master, has high accuracy, and can successfully realize efficient and accurate identification of Rubi Radix et Rhizoma original plant and congeneric similar confusable species thereof.

Description

technical field [0001] The invention relates to the technical field of identification of the original plant species of traditional Chinese medicinal materials, in particular to a method and application of using the ITS2 fragment of ribosomal DNA to identify the original species of the root of the She medicine Twisted twig and the leaves, roots and fruits of the same genus and similar easily mixed species. Background technique [0002] She medicine is a national medicine widely circulated and used among She people. It is one of the important components of traditional national medicine in my country. In 2008, She medicine was listed as "national intangible cultural heritage" by the State Council for key protection. She medicine protection , Development and utilization have a long way to go. The root of Twisted Kungfu is a traditional medicinal material of the She people. The dry root of chingii Hu is homologous to the traditional Chinese medicine raspberry (immature fruit) and...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 吕群丹程科军方洁程文亮徐金标潘俊杰
Owner LISHUI AGRI SCI
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