Comparison identifying method for dendrobium huoshanense and dendrobium hercoglossum

A technology of Dendrobium huoshanense and Dendrobium dendrobii is applied in the field of comparison and identification of Dendrobium huoshanense and Dendrobium dendrobii, which can solve problems such as unfavorable on-site detection, long time consumption, and the use of large-scale instruments.

Inactive Publication Date: 2016-11-23
WEST ANHUI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Li Dan et al. identified Dendrobium Huoshanense and its close relatives using DNA barcode ITS sequences (Li Dan, Li Zhenjian, Mao Ping, Yan Xuefeng, Chunze, Ma Xinrong, Identification and Phylogenetic Analysis of Dendrobium Materials Based on ITS Sequences, Journal of Horticultural Science, 2012, 39(8): 1539), but this method requires sequencing, which takes a long time and uses large instruments, which is not conducive to rapid and on-site detection

Method used

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  • Comparison identifying method for dendrobium huoshanense and dendrobium hercoglossum
  • Comparison identifying method for dendrobium huoshanense and dendrobium hercoglossum
  • Comparison identifying method for dendrobium huoshanense and dendrobium hercoglossum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the preparation and use method of the kit for identifying Dendrobium Huoshanense

[0025] 1. Design and synthesis of primer pairs for identification of Dendrobium huoshanense

[0026] The ITS sequences of Dendrobium Huoshanense and its obfuscated products were obtained from GenBank or sequenced in this study, as shown in Table 1 below. Among them, the haplotype sequences (including Hap5, Hap6 and Hap7) obtained by sequencing in this study are shown in sequence 4 to sequence 6 in the sequence listing.

[0027] Table 1 ITS sequences of Dendrobium Huoshanense, Dendrobium heavy lip and some similar species

[0028] Numbering

sample name

Number of samples

haplotype

GenBank

Login ID

The serial number obtained in this study

1

Dendrobium officinale Dendrobium

catenatum

1

Hap1

KP743543

2

Dendrobium officinale Dendrobium

catenatum

1

Hap2

KF143439

3

Dendrob...

Embodiment 2

[0045] Embodiment 2, using the kit prepared in Example 1 to identify the specificity analysis of Dendrobium Huoshanense

[0046] Samples to be tested: Dendrobium Huoshanense (collected from Anhui) and Dendrobium double-lipped (collected from Guangxi). All of them conformed to the description of the identification characteristics in Flora of China and related literature. Through identification, the physical objects of each sample are consistent with the names, and the quality meets the standards.

[0047] 1. Extract genomic DNA from the sample to be tested

[0048] About 50 mg of dry samples were taken respectively (of course, 100 mg of fresh samples can also be used for genomic DNA extraction), and total DNA was extracted by CTAB method. details as follows:

[0049] Put the dry medicinal material without mildew in a pulverizer to grind and pulverize, and pass through a 40-mesh sieve. Transfer the powder to a 2.0 mL microcentrifuge tube and add 900 µL of sterilized CTAB ext...

Embodiment 3

[0060] Embodiment 3, using the kit prepared in Example 1 to identify the sensitivity analysis of Dendrobium Huoshanense

[0061] Sample to be tested: Huoshan Dendrobium (collected in Anhui). Through the identification, the actual sample is consistent with the name, and the quality meets the standard.

[0062] 1. Extract genomic DNA from the sample to be tested

[0063] Take about 50 mg of dry samples (of course, 100 mg of fresh samples can also be used for genomic DNA extraction), and use CTAB method to extract total DNA. For specific operations, refer to Step 1 of Example 2.

[0064] 2. PCR amplification

[0065] The genomic DNA extracted from Dendrobium Huoshanense in Step 1 was serially diluted to obtain genomic DNA concentrations of 143 ng / μl, 71 ng / μl, 36 ng / μl, 18 ng / μl, 9 ng / μl, and 4 ng / μl, respectively. The serial dilutions of μl and 2 ng / μl were respectively used as templates, and the primer pair designed in Step 1 of Example 1 (upstream primer CP6s and downstrea...

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Abstract

The invention discloses a comparison identifying method for dendrobium huoshanense and dendrobium hercoglossum. A primer pair for identification or assistant identification of dendrobium huoshanense provided by the invention is specifically the primer pair consisting of two single-stranded DNA molecules shown by the sequence 1 and the sequence 2 in a sequence table. Experiments prove that rapid and accurate identification of dendrobium huoshanense is realized through a rapid PCR method by adopting the primers provided by the invention, and technical support is provided for the field application of molecular identification of medicinal materials.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, relates to the field of identification of traditional Chinese medicines and medicinal materials, and in particular relates to a comparison and identification method for Dendrobium Huoshanense and Dendrobium double-lipped. Background technique [0002] Huoshan Dendrobium ( Dendrobium huoshanense ) is a rare Chinese herbal medicine that is unique to the Dabie Mountains in Anhui Province and is protected by the state. It belongs to the Orchidaceae (Orchidaceae) Dendrobium ( Dendrobium ). Dendrobium heavy lip ( Dendrobium hercoglossum ) is a closely related species of Dendrobium Huoshanense. The morphological characteristics of the two are relatively similar. After making a maple bucket, the two are not easy to identify by appearance, which leads to Dendrobium double-lipped Dendrobium becoming one of the confused products of Dendrobium Huoshanense in the market. Because it is difficu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6895C12Q2531/113
Inventor 赵群陈乃富陈存武韩邦兴戴军刘枫
Owner WEST ANHUI UNIV
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