Molecular Biology and Genetics
 

Bioinformatics and Biotechnology

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 13 No. 1, Issue of January 15, 2010
© 2010 by Pontificia Universidad Católica de Valparaíso -- Chile Received September 24, 2008 / Accepted September 14, 2009
DOI: 10.2225/vol13-issue1-fulltext-13
RESEARCH ARTICLE

Molecular authentication of the traditional Chinese medicinal plant Angelica sinensis based on internal transcribed spacer of nrDNA

Tu Feng#
Laboratory of Systematic and Evolutionary Botany
College of Life Sciences
Sichuan University
Chengdu, China

Shuang Liu
Laboratory of Systematic and Evolutionary Botany
College of Life Sciences
Sichuan University
Chengdu, China

Xing-jin He*
Laboratory of Systematic and Evolutionary Botany
College of Life Sciences
Sichuan University
Chengdu, China
E-mail: xjhe@scu.edu.cn

*Corresponding author

Financial support: Nationally Natural Science Foundation of China (No. 30670146) and the National Infrastructure of Natural Resources for Science and Technology (No. 2005DKA21403).

Keywords: Angelica sinensis, internal transcribed spacer, molecular authentication, specific primers, traditional Chinese medicine.

Present address: #Department of Environment and Life sciences, Bijie University, Bijie, Guizhou, China, 551700.

Abbreviations:

AP-PCR: arbitraily primed PCR
ARMS: amplification-refractory mutation system
DMSO: dimethyl sulfoxide
GC: guanine-cytosine
ITS: internal transcribed spacer
MP: maximum parsimony
nrDNA: nuclear ribosomal DNA
RAPD: random amplified polymorphic DNA
RFLP: restriction fragment length polymorphism
SCAR: sequence characterized amplified region
TCM: traditional Chinese medicine

Abstract   Full Text

Traditionally, the authentication of the traditional Chinese medicines (TCM), Angelica sinensis, is based on slightly different morphological characters and complex compounds. Usually, those methods are simultaneously affected by several factors, leading to subtle and ambiguous results. In this study, the internal transcribed spacer (ITS) regions of A. sinensis and seven other Angelica species used as adulterants were sequenced. A pair of specific primers was designed from the polymorphic ITS regions to distinguish A. sinensis from the adulterants and regional substitutes. These ITS-derived primers amplified approximately 520 bp specific fragments from the adulterants, whereas no products was amplified with the DNA of A. sinensis. We tested eight commercially crude materials purchased in the market by using these specific primers. The result showed that there were four samples adulterating A. sinensis with regional substitutes. This indicated that A. sinensis could be accurately distinguished from the adulterants and regional substitutes. Therefore, the method of molecular authentication based on the ITS sequences may be contributed to raw material production and quality control of A. sinensis.

Supported by UNESCO / MIRCEN network