Genetic Identification of Chinese Drug Materials in Yams (Dioscorea spp.) by RAPD Analysis
HORNG-LIANG LAY1, HUNG-JEN LIU2*, MING-HUEI LIAO2, CHIEN-CHIH CHEN3, SIN-YIE LIU4 AND BOR-WEN SHEU1
1. Department of Horiculture, National Ping-Tung University of Science and Technology, 1 Hseuh-Fu Rd., Nei-Pu Hsiang, Pingtung, Taiwan, R.O.C.
2. Department of Veterinary Medicine, National Ping-Tung University of Science and Technology, 1 Hseuh-Fu Rd., Nei-Pu Hsiang, Pingtung, Taiwan, R.O.C.
3. National Research Institute of Chinese Medicine, 155-1, Sec. 2, Li-Nung St., Peitou, Taipei, Taiwan, R.O.C.
4. Executive Yuan, Council of Agriculture, Taiwan Agricultural Research Institute, 189 Chung-Cheng Rd., Wan-Feng, Wu-Feng Hsiang, Taichung, Taiwan, R.O.C.
(Received: December 19, 2000; Accepted: May 21, 2001)
ABSTRACT
The botanical origins of Chinese drug materials in yams were examined among eighteen cultivarities/ species of Dioscorea by randomly amplified polymorphic DNA (RAPD). DNA isolated from fresh leaves were used as a template in polymerase chain reactions (PCR) using thirty random decamer primers. Eighteen primers produced 237 polymorphic RAPD markers, which were scored for all the lines studied. Four primers including OPP-04, OPP-15, OPAP-04 and OPAP-10, could distinguish all tested samples. Genetic distances and similarities among eighteen cultivarities/species were calculated, and a cluster analysis was used to generate a dendrogram showing a phylogenetic relationship. The studied Dioscorea species could be divided into five subgroups by phylogenetic analysis. Genetic distance and similarity analysis showed that line 70W37 was distantly related to 70W47, and line 70W41 was closely related to 70W59. RAPD technique could be successfully applied for the rapid identification and differentiation of Dioscorea species.
Key words: botanical origins, yam (Dioscorea spp.), phylogenetic analysis