Specific PCR molecular identification of Dendrobium chrysotoxum
10.7501/j.issn.0253-2670.2013.06.021
- Author:
Si-Hao ZHENG
1
Author Information
1. Institute of Medicinal Plant Development
- Publication Type:Journal Article
- Keywords:
Dendrobium chrysotoxum Lindl.;
Dendrobium Sw.;
Molecular identification;
PCR;
Specificity
- From:
Chinese Traditional and Herbal Drugs
2013;44(6):744-748
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To design a pair of PCR primers that could identify Dendrobium chrysotoxum specifically, to optimize the system of PCR detection, and to establish a method to identify D. chrysotoxum rapidly and accurately. Methods: From GenBank database, rDNA 170 ITS sequences in the plants of Dendrobium Sw. were downloaded and compared with all sequences using MEGA 5.0; The variation sites were located, and a pair of specific primers in the non-conservative district were designed. PCR amplification was performed using the specific primers with 35 DNA templates in the plants of Dendrobium Sw., D. chrysotoxum was positive. Results: D. chrysotoxum could be specifically amplificated by specific primers when the annealing temperature was raised to 58 °C, while other plants of Dendrobium Sw. were shown as negative, and the sensitivity of the primer could reach 0.69 ng/μL. Conclusion: This study designs a method that could identify D. chrysotoxum specifically. Using this specific primer could identify D. chrysotoxum rapidly and accurately from the homologous species. This method is well-performed in specificity, and it is more simple, convenient, efficient, and accurate than other methods, such as morphological and microscopical identification, chromatograph, and spectral method.
