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Method for identifying dendrobium huoshanense by using chloroplast microsatellite primer pair

A technology of Dendrobium Huoshanense and primer pairs, which is applied to the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problems of unfavorable on-site detection, long time consumption, and the use of large instruments

Active Publication Date: 2016-11-16
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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  • Method for identifying dendrobium huoshanense by using chloroplast microsatellite primer pair
  • Method for identifying dendrobium huoshanense by using chloroplast microsatellite primer pair

Examples

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Embodiment 1

[0023] Example 1. Preparation and use method of 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 obtained from GenBank. GenBank accession number is NC_028430. The SSRIT software was used to search for the polynucleotide base repetitive sequences in the complete sequence of the chloroplast genome, and then analyzed and corrected by SciRoKo 2.1 software. A total of 52 simple repetitive sequences were searched. Use primer5 and oligo6 software to control the content (AT) 23 Design specific PCR primers for detection of Dendrobium huoshanense at both ends of simple repetitive sequence sites (chloroplast microsatellite sites), as follows:

[0026] Upstream primer CPSSR1s: 5’-CGTATGCGATAGAAGAAT-3’ (sequence 1);

[0027] Downstream primer CPSSR1a: 5'-ATCTGTAGATTGGGCTCT-3' (sequence 2).

[0028] The theoretical PCR product...

Embodiment 2

[0040] Example 2. Using the kit prepared in Example 1 to identify the specificity analysis of Dendrobium huoshanense

[0041] Samples to be tested: Dendrobium huoshanense (collected in Anhui), Dendrobium guangdong (collected in Guangxi), Dendrobium Luohe (collected in Guangxi), Dendrobium Drumstick (collected in Guangxi), Dendrobium ballflower (collected in Guangxi) ), Poly Dendrobium (collected in Guangxi), Dendrobium edulis (collected in Guangxi), Dendrobium Meihua (collected in Guangxi) and Dendrobium Huanghua (collected in Guangxi). All are in line with the description of the identification characteristics of the Flora of China. Through identification, the physical objects of each sample are consistent with the name, and the quality meets the standard.

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

[0043] Take about 50 mg of dried samples (of course, 100 mg of fresh samples can also be used for genomic DNA extraction), and extract total DNA by CTAB method. deta...

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Abstract

The invention discloses a method for identifying dendrobium huoshanense by using a chloroplast microsatellite primer pair. The primer pair used for identifying dendrobium huoshanense or assisting identification of dendrobium huoshanense is a primer pair consisting of two single-chain DNA molecules as shown in sequence 1 and sequence 2 in a sequence table. Experiments prove that, by adopting the primer, the dendrobium huoshanense can be quickly and accurately identified from dendrobium wilsonii, dendrobium lohohense, dendrobium chrysotoxum, dendrobium thyrsiflorum, dendrobium lindleyi, dendrobium denndanum, dendrobium loddigesii, dendrobium dixanthum and other adulterants by adopting a quick PCR method, so that technical support is provided to field application of medicinal material molecular identification.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology, relates to the field of Chinese medicine and Chinese medicinal material identification, in particular to a chloroplast microsatellite primer pair for identifying Dendrobium huoshanense and its application. Background technique [0002] Dendrobium huoshanense ( Dendrobium huoshanense ) Is a rare Chinese herbal medicine that is unique to the Dabie Mountains in Anhui Province and is under national key protection. It belongs to the Orchidaceae (Orchidaceae) Dendrobium ( Dendrobium ). Because there are more relative plants of the same genus of Dendrobium Huoshan, there are more confusing products. Some obfuscated products currently on the market include Dendrobium Guangdong, Dendrobium Luohe, Dendrobium Dendrobium, Dendrobium ballflower, Dendrobium Polydendrum, Dendrobium Dioscorea, Dendrobium Meihua, Dendrobium Huanghua, etc. [0003] The morphological characteristics of Dendrobium huoshanense a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
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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