DNA molecular identification of Chinese herbal materials of Dysosma by ITS2 barcode sequence / 中草药

ABSTRACT

Objective To explore a new identification method for medicinal materials of Dysosma, and analyze the second internal transcribed spacer (ITS2) barcode sequences of Diphylleia sinensis and Dysosma versipellis, Sinopodophyllum hexandrum, Dysosma difformis and Dysosma pleiantha in five kinds of podophyllum. Methods The ITS2 of ribosomal DNA of medicinal materials of podophyllum was amplified and sequenced by bi-directional sequencing of PCR products. Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner. Phylogenetic study was performed using software MEGA 5.1 in accordance with Kimura-2-parameter (K2P) model. Genetic distances were calculated and analyzed and the phylogenetic tree was constructed by using the neighbor-joining (NJ) method. Results There were significant differences among five kinds of Dysosma. Their maximum intraspecific genetic distance (K2P distance) was far lower than their minimum interspecific genetic distance with the other species. In the cluster dendrogram, all species showed monophyletic. Conclusion ITS2 sequence as DNA barcoding technique can be used to identify Chinese herbal materials of Dysosma.