Molecular Differentiation of Atractylodes Drugs by PCR-Restriction Fragment Length Polymorphism and PCR-Selective Restriction Analysis on the 18S-5.8S rDNA Intratranscribed Spacer 1 Gene

Hwei-Fang Cheng*, Been Lai, Shiu-Chen Chan, Ching-Pang Chou, Te-Hsiun Yang, Wen-Hong Huang, Chun-Heng Liao and Chia-Po Lin

National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Taipei, Taiwan, R. O. C.

ABSTRACT

   The 18S-5.8S rDNA intratranscri bed spacer 1 (ITS1) regions from Atractylodes japonica, A. lancea and A. ovata rhizomes were amplified using the polymerase chain reaction (PCR) with consensus rRNA gene primers. The amplified DNA fragments from Atractylodes species were of similar size, but with different MspI restriction mapping. Cloning and sequencing of the PCR products showed that the DNA sequences of PCR-amplified ITS1 were distinct between Atractylodes species. Sequence analysis indicated that the amplified ITS1 elements were informative for ITS1 fingerprinting following digestion with 6-cutter restriction endonucleases BspEI, AflII, and NaeI. Therefore, both polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-selective restriction (PCR-SR) analysis on the ITS1 gene were established to provide useful tools for the authentication between Atractylodes rhizomes.

Key words: Atractylodes japonica, Atractylodes lancea, Atractylodes ovata, ITS1, PCR-RFLP, PCR-SR, herbal origin