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DNA barcode identification method of traditional She medicinal Shiliang herb tea based original species and related confusable species

A barcode and herbal tea technology, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the controversies in the taxonomy, origin and evolution of Wintersweet plants, few studies on the identification of the original species, identification difficulties, etc. problem, to achieve the effect of easy grasp, high accuracy and wide applicability

Inactive Publication Date: 2017-06-13
LISHUI AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The morphology of Chimeras plants is very close, and traditional morphological identification is difficult, which leads to disputes in the systematic classification, origin and evolution of Chimeras plants. Therefore, misuse and mixed use of close relatives in actual production and clinical applications are sometimes easy to mix. occur
At present, most of the studies on herbal teas focus on pharmacology and chemical components, but there are few studies on the identification of its basic species. Only Zhu Yeqing et al. have systematically classified and identified Wintersweet from the aspects of traditional morphology. Species identification using modern molecular biology techniques

Method used

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  • DNA barcode identification method of traditional She medicinal Shiliang herb tea based original species and related confusable species
  • DNA barcode identification method of traditional She medicinal Shiliang herb tea based original species and related confusable species
  • DNA barcode identification method of traditional She medicinal Shiliang herb tea based original species and related confusable species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 Distinguishes edible herbal tea base plant and Wintersweet, Wintersweet are easy to mix species

[0029] 1 provide samples

[0030] A total of 37 samples of herbal tea base plants and leaves of the same genus and easily mixed species were collected from different origins (see Table 1 for details) Table 1 Sample information table

[0031]

[0032]

[0033] 2 Genomic DNA Extraction

[0034] 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 extraction kit (Tiangen Biotech Co., China) , in a 65°C water bath for 1 h, and the rest of the steps were carried out according to the instructions of the kit.

[0035] 3PCR amplification

[0036] In the PCR reaction system

[0037] Forward primer: SEQ ID No.1: 5′-ATGCGATACTTGGTGTGAAT-3′;

[0038] Reverse primer: SEQ ID No.2: 5′-GACGCTTCTCCAGACTACAAT-...

Embodiment 2

[0052] Embodiment 2 Differentiate edible herbal tea base plant and the same genus easily mixed species

[0053] 1 provide samples

[0054] A total of 37 samples of herbal tea base plants and leaves of the same genus and easily mixed species were collected from different origins (see Table 2 for details) Table 2 Sample information table

[0055]

[0056]

[0057] 2 Genomic DNA Extraction

[0058] 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 extraction kit (Tiangen Biotech Co., China) , in a 65°C water bath for 1 h, and the rest of the steps were carried out according to the instructions of the kit.

[0059] 3 sequencing

[0060] In the PCR reaction system

[0061] Forward primer: SEQ ID No.1: 5′-ATGCGATACTTGGTGTGAAT-3′;

[0062] Reverse primer: SEQ ID No.2: 5′-GACGCTTCTCCAGACTACAAT-3′;

[0063] React...

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Abstract

The invention relates to the field of Chinese herbal medicine based original plant species identification technology, specifically to a method for identifying traditional She medicinal Shiliang herb tea based original species and related confusable species by the utilization of ITS2 fragments of ribosomal DNA and application thereof. A DNA barcode identification method of traditional She medicinal Shiliang herb tea based original species and related confusable species comprises the following steps: 1) DNA sample extraction; 2) PCR amplification of fragments containing ITS2 sequences; 3) splicing of PCR amplification products; 4) comparison of ITS2 sequences of multiple species; and 5) construction of phylogenetic tree based on the ITS2 sequences. The DNA extraction method of traditional She medicinal Shiliang herb tea based original species and related confusable species and the ITS2 preparation method can effectively solve the problems such as species identification of Shiliang herb tea and related species, development and utilization of germplasm resources, etc. The method of the invention has extensive applicability, is simple to operate, is easy to grasp and has high accuracy. By the method, rapid and accurate identification of the Shiliang herb tea based original species and related confusable species can be successfully realized.

Description

technical field [0001] The invention relates to the technical field of identification of 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 herbal tea and its closely related and easily mixed species. Background technique [0002] She medicine is an important part of the motherland's treasure house of medicine and a part of the world's excellent cultural heritage. It is the product of the She people's long-term struggle against diseases in order to survive and reproduce under specific historical conditions and special geographical environments. In the process of summarizing the experience of disease prevention and treatment for many years, the medicine with national characteristics was gradually formed. As a unique medicinal material of the She nationality, herbal tea has a long history of folk use. Its base species are the dried leaves of Chimonanthus salicifoli...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156C12Q2600/172
Inventor 吕群丹程科军姚辉马双姣程文亮方洁徐金标姜程曦周红
Owner LISHUI AGRI SCI
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