Identification of the Crude Drug Atractylodes Rhizome (Byaku-jutsu) and Atractylodes Lancea Rhizome (So-jutsu) Using Chloroplast TrnK Sequence as a Molecular Marker

Novel methods for molecular authenticaiton of Atractylodes-derived crude drugs (Jutsu) were established based on PCR-restriction fragment length polymorphism (RFLP) and direct sequencing of chloroplast trnK. Two regions inside the chloroplast trnK were selected as molecular markers for identificatio...

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Bibliographic Details
Published in:Biological & pharmaceutical bulletin 2000/05/01, Vol.23(5), pp.589-594
Main Authors: MIZUKAMI, Hajime, OKABE, Yuka, KOHDA, Hiroshi, HIRAOKA, Noboru
Format: Article
Language:English
Subjects:
Asteraceae - chemistry
Asteraceae - genetics
Atractylodes Lancea Rhizome
Atractylodes Rhizome
Biological and medical sciences
chloroplast trnK
Chloroplasts - genetics
crude drug authentication
direct sequencing
DNA, Plant - analysis
Drugs, Chinese Herbal - analysis
General pharmacology
Genes, Plant
Genetic Markers
Medical sciences
Molecular Sequence Data
PCR-RFLP
Pharmacognosy. Homeopathy. Health food
Pharmacology. Drug treatments
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Sequence Analysis
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Summary:Novel methods for molecular authenticaiton of Atractylodes-derived crude drugs (Jutsu) were established based on PCR-restriction fragment length polymorphism (RFLP) and direct sequencing of chloroplast trnK. Two regions inside the chloroplast trnK were selected as molecular markers for identification and discrimination of Atractylodes Rhizome (Byaku-jutsu) and Atractylodes Lancea Rhizome (So-jutsu). The Region 1 fragment (260 bp) amplified from So-jutsu and Wa-byaku-jutsu (Atractylodes Rhizome derived from A. japonica) gave 2 bands of 180 bp and 80 bp on agarose gel electrophoresis after digestion with a restriction endonuclease HinfI, whereas the fragment amplified from Kara-byaku-jutsu (Atractylodes Rhizome derived from A. ovata) remained undigested, which allowed unambiguous identification of Kara-byaku-jutsu. By direct sequencing of Region 2 (436 bp) and comparison of the nucleotide sequence data sets we could not only discriminate Byaku-jutsu and So-jutsu but also identify the original plant species of each crude drug specimen. A simple and reliable protocol for rapid preparation of DNA suitable for PCR from as little as 1 mg of Atractylodes-derived crude drugs was also described.
ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.23.589