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A molecular identification method and application of Dendrobium candidum dry medicinal material

A technology of Dendrobium officinale and medicinal materials, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA preparation, etc., can solve the problems of long time, low amplification success rate, sample DNA degradation, etc., and achieve high accuracy, Widely applicable, easy-to-master effects

Active Publication Date: 2019-06-04
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, this technology has the following problems in the identification of Dendrobium candidum dry medicinal materials: (1) During the processing of Tiepi Fengdou, there is usually a heating process at 80°C to 100°C, and the time is long, which will cause serious degradation of sample DNA and affect DNA extraction efficiency; (2) the polysaccharide content of Dendrobium officinale is high, which seriously affects the extraction and purification of DNA; (3) the universal primer ITS2F / 3R has a low amplification success rate in the sequence amplification of the dry medicinal material ITS2 of Dendrobium officinale

Method used

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  • A molecular identification method and application of Dendrobium candidum dry medicinal material
  • A molecular identification method and application of Dendrobium candidum dry medicinal material
  • A molecular identification method and application of Dendrobium candidum dry medicinal material

Examples

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Embodiment 1

[0020] The identification of embodiment 1 Dendrobium candidum medical material

[0021] 1. Experimental materials

[0022] In this case, a total of 173 samples of 40 species of Dendrobium were collected, and 3 samples of fake Dendrobium dendrobii and Dendrobium fringe were collected. Among them, there are 14 original plant samples of Tiepi Fengdouji and 65 samples of medicinal materials. The specific information is shown in Table 1.

[0023] Table 1 Sample information table of Tiepi Fengdou and its counterfeit products

[0024]

[0025]

[0026]

[0027] 2. Experimental equipment

[0028] Desktop high-speed centrifuge, PTC-100 gene amplification instrument, electrophoresis instrument, ultraviolet gel imaging analysis system, MM400 ball mill (Germany Retsch).

[0029] 3. DNA extraction

[0030] Take 20-30 mg of dried Tiepi Fengdou herbs, chop them up and crush them with MM400 ball mill, add 500-1000 μL of nuclear separation solution, mix well, centrifuge at 7000 r / ...

Embodiment 2

[0042] The identification of embodiment 2 Dendrobium officinale base plant

[0043] Basically the same as Example 1, except that the original plant of Dendrobium candidum was used as the sample, DNA was extracted according to the above method, and the ITS2 sequence was amplified. As a result, the target sequence was also successfully obtained and the identification was completed.

Embodiment 3

[0044] The identification of embodiment 3 dendrobium officinale seed seedlings

[0045] Basically the same as Example 1, the difference is that Dendrobium officinale seeds and seedlings are used as samples, DNA is extracted according to the above method, and the ITS2 sequence is amplified. As a result, the target sequence can also be successfully obtained and the identification is completed.

[0046]

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Abstract

The invention discloses a molecular identification method of a rare traditional Chinese medicine dry dendrobium officinale medicinal material and application thereof, and can effectively solve the rapid identification problem of dry dendrobium officinale medicinal material. The specific steps include: (1) preparing a to-be-detected sample DNA template by improved DNA extraction method; (2) amplifying the sample rDNA ITS2 fragment through specific PCR amplification primers and reaction parameters; (3) sequencing the amplification product to obtain an ITS2 sequence of the dry dendrobium officinale medicinal material; and (4) constructing an ITS2 sequence NJ tree of the sample. The method provided by the invention can realize rapid and accurate identification of dry dendrobium officinale medicinal material, original plants, and seeds and seedlings, etc.

Description

technical field [0001] The invention relates to the technical field of identification of source varieties of terminology Chinese medicinal materials, in particular to a method for rapidly identifying dry medicinal materials of Dendrobium candidum by using ribosomal DNA ITS2 fragments and an application thereof. Background technique [0002] Dendrobium officinale Kimura et Migo (the Latin name Dendrobium catenatum Lindley in the English version of "Flora of China" eflora) is a rare and precious medicinal plant of Orchidaceae, and has the reputation of "the first of the nine Chinese immortal grasses". Dendrobium officinale has strict requirements on the growth environment. Its natural reproductive capacity is low, its growth is slow, and after long-term excavation, its natural resources are on the verge of exhaustion. It is listed as a national key protected medicinal material. The description of Dendrobium officinale in the 2010 edition of "Chinese Pharmacopoeia" is "This pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/10
CPCC12Q1/6806C12Q1/6895
Inventor 姚辉杨培韩建萍陈士林宋经元杨兵勋马双姣
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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