Identification of Fritillaria pallidiflora using diagnostic PCR and PCR-RFLP based on nuclear ribosomal DNA internal transcribed spacer sequences.
Fritillaria pallidiflora Schrenk (Liliaceae) is a commonly used antitussive herb. There are 9 species of Fritillaria recorded as herbal drugs in the Chinese Pharmacopoeia. The other species are often marketed as F. pallidiflora, and thus, the therapeutic effects of F. pallidiflora are not achieved. Methods to distinguish F. pallidiflora from the 8 other species of Fritillaria are limited by the current morphological and chemical methods. In this study, we report two molecular authentication methods based on the sequences of nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) regions. For diagnostic PCR, we designed a pair of species-specific primers to authenticate F. pallidiflora. The PCR program consisted of only two steps for every repeated cycle. For PCR-RFLP, we identified a distinctive site which can be recognized by the restriction endonuclease Eco81I in the nrDNA ITS1 region of F. pallidiflora. PCR-RFLP analysis was established to differentiate F. pallidiflora from the other species of Fritillaria. These methods provide effective and accurate identification of F. pallidiflora.
Wang
Chong-Zhi
CZ
Key Laboratory of Modern Traditional Chinese Medicines, and Department of Pharmacognosy, School of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, Jiangsu Province, PR China.
Li
Ping
P
Ding
Jia-Yi
JY
Jin
Guo-Qian
GQ
Yuan
Chun-Su
CS
D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis*; D021903:DNA, Ribosomal Spacer / Q000032:analysis*; D031410:Fritillaria / Q000235:genetics*; D006801:Humans; D008517:Phytotherapy; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
D017931:DNA Primers; D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer