Method of identifying the species of plant belonging to the genus uncaria

A technology of Uncaria, plants, applied in the field of identifying species of Uncaria plants

Active Publication Date: 2010-07-07
TSUMURA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many species of Uncaria plants have not yet been classified by determining their nucleotide sequences and then distinguishing species from them.

Method used

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  • Method of identifying the species of plant belonging to the genus uncaria
  • Method of identifying the species of plant belonging to the genus uncaria
  • Method of identifying the species of plant belonging to the genus uncaria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Material

[0091] Use wax leaf specimens that can be identified according to the morphology to study the variation sites that are conducive to identification. The samples used for the study are shown in Table 3. Except for Uncaria homomella, for each species, the base sequences of the ITS region were determined for a plurality of samples, and the species-specific site was studied.

[0092] table 3

[0093]

[0094] method

[0095] About 200 mg of leaves or pericarp were collected from the samples, and the total DNA was extracted using DNeasy (registered trademark) PlantMini Kit (Qiagen). Regarding the PCR amplification of the ITS region, since the amplifiable length varies depending on the state of the sample, the amplification is performed in the following procedure. Also, for samples that are difficult to amplify, try to purify the entire DNA using the Plasmid Mini Kit (Qiagen).

[0096] (1) Using a primer set (ITS5 & ITS4), ITS1, 5.8S, and ITS2 were ampli...

Embodiment 2

[0116] By analyzing the nucleotide sequences of the ITS region of Diaoyinghua (crude drug) obtained from Huangjing Township, Gulin County, Sichuan Province, and Diaoyinghua (crude drug) obtained from a production company in Sichuan Province, it was identified which Uncaria plant was included.

[0117] Material

[0118] As samples, 2 samples of rattan hooks obtained from Huangjing Township, Gulin County, Sichuan Province and 2 batches of 8 samples of rattan hooks obtained from Chengdu Longquan Natural Medicine Development Co., Ltd. were used.

[0119] method

[0120] Total DNA was extracted from about 200 µg of the sample using DNeasy (registered trademark) Plant Mini Kit (Qiagen). In the PCR amplification of the ITS region, primer sets (ITS5 & ITS3) were used. The composition of the reaction solution is as follows: 5 μl of 10×Buffer, 4 μl of dNTP mix (2.5 mM), 2.5 μl×2 of primer (10 pmol / μl), 30.75 μl of 10% DMSO, 5 μl of template DNA, and 0.25 μl of Gene-Taq. The react...

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Abstract

It is intended to provide a method whereby the species of a plant belonging to the genus Uncaria can be identified at a high accuracy. Namely, a method of identifying the species of a plant belonging to the genus Uncaria contained in a test sample which comprises comparing the base sequence in the ITS region of the ribosomal DNA of the plant contained in the test sample with at least one base sequence in the ITS region selected from among the base sequences represented by SEQ ID NOS:1 to 11.

Description

technical field [0001] The present invention relates to a method for identifying species of Uncaria plants by genetic methods. Background technique [0002] Fishing hook is a crude drug recorded in the 14th edition of the Japanese Pharmacopoeia, Supplement 1. According to its records, Uncaria rhynchophylla Miquel, Uncaria sinensis Haviland, and Uncaria macrophylla Wallich Three species are the original plants. Yet, there are 11 kinds of Uncaria plants (Hsue H. & Wu H., 1999.Uncaria in LoH.ed.Flora Reipublicae Popularis Sinicae 71 (1) Science Press, Beijing) that are distributed in China, and there are also circulations derived from the above-mentioned 3 Herbal medicines of species other than this species. However, it is difficult to distinguish such a crude drug from a true crude drug containing the three species of Uncaria genus. Therefore, as the traditional Chinese medicine preparation of fishing rattan hook, there are problems in its quality stability. Regarding this...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/09C12Q1/6895
CPCC12Q1/6895A01H5/02Y10T436/143333
Inventor 山路弘树
Owner TSUMURA
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