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Method for identifying dendrobium huoshanense and dendrobium henanense through comparison

A Dendrobium Huoshanense, identification method technology, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of unfavorable on-site detection, use of large instruments, and long time consumption, etc., and achieve low requirements for equipment and time-consuming Short, high-sensitivity effects

Inactive Publication Date: 2017-01-04
WEST ANHUI UNIV
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  • 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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  • Method for identifying dendrobium huoshanense and dendrobium henanense through comparison
  • Method for identifying dendrobium huoshanense and dendrobium henanense through comparison
  • Method for identifying dendrobium huoshanense and dendrobium henanense through comparison

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the preparation and use method of the kit for identifying Dendrobium Huoshanense

[0029] 1. Design and synthesis of primer pairs for identification of Dendrobium huoshanense

[0030] 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.

[0031] Table 1 ITS sequences of Dendrobium Huoshanense, Dendrobium Henan and some similar species

[0032] serial number sample name Number of samples haplotype GenBank accession number The serial number obtained in this study 1 Dendrobium officinale Dendrobium catenatum 1 Hap1 KP743543 2 Dendrobium officinale Dendrobium catenatum 1 Hap2 KF143439 3 Dendrobium nobile Dendrobium nobile 1 Hap3 EF61...

Embodiment 2

[0049] Embodiment 2, using the kit prepared in Example 1 to identify the specificity analysis of Dendrobium Huoshanense

[0050] Samples to be tested: Dendrobium Huoshanense (collected from Huoshan, Anhui) and Henan Dendrobium (collected from Huoshan, Anhui). 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.

[0051] 1. Extract genomic DNA from the sample to be tested

[0052] 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:

[0053] 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 ...

Embodiment 3

[0064] Embodiment 3, using the kit prepared in Example 1 to identify the sensitivity analysis of Dendrobium Huoshanense

[0065] Sample to be tested: Huoshan Dendrobium (collected from Huoshan, Anhui). Through the identification, the actual sample is consistent with the name, and the quality meets the standard.

[0066] 1. Extract genomic DNA from the sample to be tested

[0067] 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.

[0068] 2. PCR amplification

[0069] The genomic DNA extracted from Dendrobium Huoshanense in Step 1 was serially diluted to obtain genomic DNA concentrations of 156 ng / μl, 78 ng / μl, 39 ng / μl, 19 ng / μl, 10 ng / μl, and 5 ng / μl, respectively. The serial dilutions of μl and 2 ng / μl were respectively used as templates, and the primer pair (primers SH-CP29s and SH-CP29a) designed in Step 1 of E...

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Abstract

The invention relates to a method for identifying dendrobium huoshanense and dendrobium henanense through comparison. The method comprises comparison identification operation steps as follows: (1) genomic DNA is extracted from two to-be-detected samples and taken as a template, primers are adopted to perform a PCR (polymerase chain reaction) amplification reaction respectively, and two PCR amplified products are obtained; (2) the two PCR amplified products are detected with an agarose gel electrophoresis method, and if one of the PCR amplified products contains DNA fragments of 262bp, one of the to-be-detected samples contains the dendrobium huoshanense or a candidate contains the dendrobium huoshanense; if the PCR products don't contain the DNA fragments of 262bp, one of the to-be-detected samples doesn't contain the dendrobium huoshanense or a candidate doesn't contain the dendrobium huoshanense. By means of the method, the dendrobium huoshanense and the dendrobium henanense are rapidly and accurately identified.

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, in particular to a method for identifying Dendrobium Huoshanense and Dendrobium Henan. 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 ). Henan Dendrobium ( Dendrobium henanense ) is a closely related species of Dendrobium Huoshanense, and the morphological characteristics of the two are relatively similar, especially after they are made into maple buckets, it is more difficult to identify the two through their appearance, resulting in Henan Dendrobium becoming one of the confused products of Dendrobium Huoshanense in the market. Due to the difficulty in identifying Dendr...

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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