A method for identifying Dendrobium Huoshanense with chloroplast microsatellite primer pairs

A technology of Dendrobium Huoshanense and primer pairs, which is applied in the direction of biochemical equipment and methods, and microbial measurement/inspection, and can solve problems such as unfavorable on-site detection, use of large instruments, and long time consumption

Active Publication Date: 2019-05-28
WEST ANHUI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A method for identifying Dendrobium Huoshanense with chloroplast microsatellite primer pairs
  • A method for identifying Dendrobium Huoshanense with chloroplast microsatellite primer pairs

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Experimental program
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Embodiment 1

[0023] Embodiment 1, the preparation and use method of the kit for identifying Dendrobium Huoshanense

[0024] 1. Design and synthesis of primer pairs for identification of Dendrobium huoshanense

[0025] The complete sequence of the chloroplast genome of Dendrobium Huoshanense was searched from GenBank, and the GenBank accession number is NC_028430. SSRIT software was used to search for polynucleotide base repeat sequences in the complete sequence of the chloroplast genome, and then analyzed and corrected by SciRoKo 2.1 software. A total of 52 simple repeat sequence sites were searched. Use primer5 and oligo6 software to contain (AT) 23 The two ends of the simple repeat sequence site (chloroplast microsatellite site) were designed to detect specific PCR primers for Dendrobium huoshanense, as follows:

[0026] Upstream primer CPSSR1s: 5'-CGTATGCGATAGAAGAAT-3' (SEQ ID NO: 1);

[0027] Downstream primer CPSSR1a: 5'-ATCTGTAGATTGGGCTCT-3' (SEQ ID NO: 2).

[0028] Theoretical P...

Embodiment 2

[0040] Embodiment 2, using the kit prepared in Example 1 to identify the specificity analysis of Dendrobium Huoshanense

[0041] Samples to be tested: Huoshan Dendrobium (collected from Anhui), Guangdong Dendrobium (collected from Guangxi), Luohe Dendrobium (collected from Guangxi), Drumstick Dendrobium (collected from Guangxi), Ball Flower Dendrobium (collected from Guangxi ), Dendrobium poly(collected from Guangxi), Dendrobium multiflora (collected from Guangxi), Dendrobium meihua (collected from Guangxi) and Dendrobium chrysanthemum (collected from Guangxi). All of them conformed to the description of the identification characters of Flora of China. Through identification, the physical objects of each sample are consistent with the names, and the quality meets the standards.

[0042] 1. Extract genomic DNA from the sample to be tested

[0043] About 50 mg of dry samples were taken respectively (of course, 100 mg of fresh samples can also be used for genomic DNA extraction...

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Abstract

The invention discloses a method for identifying Dendrobium Huoshanense with chloroplast microsatellite primer pairs. The primer pair for identifying or assisting in identifying Dendrobium Huoshanense provided by the present invention is specifically a primer pair composed of two single-stranded DNA molecules shown in sequence 1 and sequence 2 in the sequence listing. Experiments have proved that the primers provided by the present invention have been used to realize the confusion of Dendrobium Huoshanense and Dendrobium Guangdong, Dendrobium Luohe, Dendrobium chrysanthemum, Dendrobium chinensis, Dendrobium polythene, Dendrobium pleura, Dendrobium meihua and Dendrobium chrysanthemum by the rapid PCR method. Provide technical support for the rapid and accurate identification of medicinal products and the on-site 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, in particular to a pair of chloroplast microsatellite primers for identifying Dendrobium Huoshanense and its application. 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 ). Since there are many related plants of the same genus of Dendrobium Huoshanense, there are many confusions. Some confusing products currently on the market include Guangdong Dendrobium, Luohe Dendrobium, Drumstick Dendrobium, Ball Flower Dendrobium, Poly Dendrobium, Multi-sheath Dendrobium, Meihua Dendrobium and Yellow-flowered Dendrobium, etc. [0003] The morphological characteristics of Dendrobium Huos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 赵群陈乃富陈存武韩邦兴戴军刘枫
Owner WEST ANHUI UNIV
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