Dong Xiao-Man, Jiang Chao, Yuan Yuan, Zha Liang-Ping, Peng Dai-Yin, Zhao Yu-Yang
School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China.
State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2017 Mar;42(5):896-901. doi: 10.19540/j.cnki.cjcmm.20170217.001.
Based on rDNA ITS sequences of Dendrobium officinale and the other 69 species of Dendrobium, a pair of dismatched allele-specific diagnostic primers, TPSH-AS1F and TPSH-AS1R were designed to authenticate D. officinale from the other species. Thebidirectional PCR amplification were performed using the diagnostic primers with the total DNAs of the original plants or processing products as a template. When the annealing temperature was raised to 60 ℃, only the template DNA of D. officinale could be amplified whereas the diagnostic PCRs of the other Dendrobium species were all negative. Compared with the other authentification methods, the bidirectional PCR amplifications is not only simpler and time-saving but practical and effective.
基于铁皮石斛及其他69种石斛的核糖体DNA内转录间隔区(rDNA ITS)序列,设计了一对不匹配的等位基因特异性诊断引物TPSH-AS1F和TPSH-AS1R,用于鉴别铁皮石斛与其他物种。以原植物或加工产品的总DNA为模板,使用诊断引物进行双向PCR扩增。当退火温度提高到60℃时,仅能扩增出铁皮石斛的模板DNA,而其他石斛物种的诊断性PCR均为阴性。与其他鉴定方法相比,双向PCR扩增不仅更简单、省时,而且实用有效。