Planta Med 1999; 65(4): 360-364
DOI: 10.1055/s-1999-14003
Original Paper

Georg Thieme Verlag Stuttgart · New York

Molecular Diversity of 5S-rRNA Spacer Domain in Fritillaria Species Revealed by PCR Analysis

Z. H. Cai1,2 , P.  Li2 , T. T.X. Dong1 , K. W.K. Tsim1
  • 1Department of Biology and Biotechnology Research Institute, The Hong Kong University of Science and Technology, Hong Kong, China
  • 2China Pharmaceutical University, Ji Xiang An, Nanjing, China
Further Information

Publication History

August 24, 1998

February 21, 1999

Publication Date:
31 December 1999 (online)

Abstract:

Beimu (bulbs of Fritillaria) is an important traditional Chinese herbal medicine commonly used as an antitussive and expectorant. There are about 25 species and varieties of Fritillaria that carry the name Beimu on commercial markets. The price for each Beimu may differ by more than 100-fold. However, the identification of the origin of a particular species on the market is difficult. Here, a molecular method was used to identify various species of Fritillaria regardless of their geographical origin. The 5S-rRNA coding sequence is highly conserved in higher eukaryotes, but the spacer sequence of the 5S-rRNA gene is variable among different species. Total genomic DNA from fresh leaves and bulbs of Fritillaria cirrhosa, F. puqiensis, F. anhuiensis, and F. thunbergii was extracted. The 5S-rRNA spacer region of the extracted DNAs was amplified by PCR with a pair of primers located within the conserved coding region. The isolated cDNA clones (∼ 600 bp) covering the 5S-rRNA spacer domain were sequenced. By aligning the isolated nucleotide sequences of the four Fritillaria species, sequence diversity was found in the spacer region. Furthermore, a unique EcoR I site was used for the rapid identification of different species of Fritillaria. To our knowledge, this is the first report on the detection of 5S-rRNA spacer domain sequences of Fritillaria and their use to identify species.