Identification of constituent herbs in ginseng decoctions by DNA markers

Chinese Medicine

Pang-Chui Shaw²  Ming Li²  Yat-Tung Lo¹ 

[1] School of Life Sciences, The Chinese University of Hong Kong, S.A.R N.T, Shatin, Hong Kong, China;State Key Laboratory of Phytochemistry and Plant Resources in West China (CUHK), Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

关键词: DNA sequencing;    Multiplex PCR;    DNA marker;    Molecular authentication;    Decoction;    Ginseng;   

Others  :  1139130
DOI  :  10.1186/s13020-015-0029-x

 received in 2014-08-29, accepted in 2015-01-20,  发布年份 2015

【 摘 要 】

Background

DNA in herbal decoctions is usually fragmented by extensive boiling and is usually regarded as unsuitable for molecular authentication. This study aims to evaluate the feasibility for molecular authentication methods by multiplex polymerase chain reaction (PCR) and DNA sequencing for decoctions.

Methods

The DNA extraction procedure, sample pulverization and boiling time were examined in (1) single herb decoction with Panax ginseng (“ginseng”) or P. quinquefolius (“American ginseng”), (2) decoctions of two classical ginseng prescriptions of five herbal materials (Aconitum carmichaeli, Atractylodes macrocephala, P. ginseng, Glycyrrhiza uralensis and Zingiber officinale) and (3) a commercial ready-to-serve ginseng soup. Primers were designed from 26S-18S, ITS2 and trnH-psbA region, with DNA sequences obtained from GenBank. Multiplex PCR was also employed in ginseng or American ginseng decoctions to differentiate between these two herbs.

Results

All six herbal species tested in this study could be identified in decoctions. We had four main observations: (1) sample pulverization before boiling improved PCR detection; (2) prolonged boiling increased the DNA concentration but decreased the intactness of DNA fragments; (3) ginseng could be differentiated from American ginseng by multiplex PCR; (4) identification of individual herbs in multi-herb decoctions with prolonged boiling time of 180 min could be achieved.

Conclusions

DNA could be amplified from extensively boiled ginseng decoctions, multi-herb decoctions and commercial soup although DNA degradation was critical to successful PCR.

【 授权许可】

   
2015 Lo et al.; licensee BioMed Central.

【 图 表 】

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