Abstract
DNA barcoding is a novel technology that uses a standard DNA sequence to facilitate species identification. Although a consensus has not been reached regarding which DNA sequences can be used as the best plant barcodes, the psbA-trnH spacer region has been tested extensively in recent years. In this study, we hypothesize that the psbA-trnH spacer regions are also effective barcodes for Dendrobium species. We have sequenced the chloroplast psbA-trnH intergenic spacers of 17 Dendrobium species to test this hypothesis. The sequences were found to be significantly different from those of other species, with percentages of variation ranging from 0.3 % to 2.3 % and an average of 1.2 %. In contrast, the intraspecific variation among the Dendrobium species studied ranged from 0 % to 0.1 %. The sequence difference between the psbA-trnH sequences of 17 Dendrobium species and one Bulbophyllum odoratissimum ranged from 2.0 % to 3.1 %, with an average of 2.5 %. Our results support the notion that the psbA-trnH intergenic spacer region could be used as a barcode to distinguish various Dendrobium species and to differentiate Dendrobium species from other adulterating species.
Key words
Dendrobium
- Orchidaceae - DNA barcoding -
psbA‐trnH intergenic region
References
-
1
Hebert P DN, Ratnasingham S, deWaard J R.
Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species.
Proc R Soc Lond B Biol Sci.
2003;
270 (Suppl.)
S96-S99
-
2
Hebert P DN, Cywinska A, Ball S L, De Waard J R.
Biological identification through DNA barcodes.
Proc R Soc Lond B Biol Sci.
2003;
270
313-321
-
3
Kress W J, Wurdack K J, Zimmer E A, Weigt L A, Janzen D H.
Use of DNA barcodes to identify flowering plants.
Proc Natl Acad Sci USA.
2005;
102
8369-8374
-
4
Chase M W, Cowan R S, Hollingsworth P M, van den Berg C, Madriñán S, Petersen G, Seberg O, Jørgsensen T, Cameron K M, Carine M, Pedersen N, Hedderson T, Conrad F, Salazar G A, Richardson J E, Hollingsworth M L, Barraclough T G, Kelly L, Wilkinson M.
A proposal for a standardised protocol to barcode all land plants.
Taxon.
2007;
56
295-299
-
5
Shaw J, Lickey E B, Schilling E E, Small R L.
Comparision of whole chloroplast genome sequences to choose noncoding regions for phylogenetic studies in angiosperms: The tortoise and the hare III.
Am J Bot.
2007;
94
275-288
-
6
Newmaster S G, Fazekas A J, Steeves R AD, Janovec J.
Testing candidate plant barcode regions in the Myristicaceae.
Mol Ecol Resour.
2008;
8
480-490
-
7
Sucher N J, Carles M C.
Genome-based approaches to the authentication of medicinal plants.
Planta Med.
2008;
74
603-623
-
8 The State Pharmacopoeia Commission of the PRC .Pharmacopoeia of the People's Republic of China. Beijing; Chemical Industry Press 2005
-
9
Tsi Zhanhe.
The flora sinica.
Science Press of China.
1999;
19
67-146
-
10
Li M F, Xu G J, Xu L S, Jin R L.
Survey and identification of commercial example of Shihu (Dendrobium) (I).
Chin Tradit Herbal Drugs.
1986;
17
33-37
-
11
Li M F, Xu G J, Xu L S, Jin R L.
Survey and identification of commercial example of Shihu (Dendrobium) (II).
Chin Tradit Herbal Drugs.
1991;
22
173-178
-
12
Ma G X, Xu G J, Xu L S, Li M F.
Survey and identification of commercial example of Shihu (Dendrobium) (III).
Chin Tradit Herbal Drugs.
1995;
26
370-372
-
13
Stern W L, Morris M W, Judd W S.
Anatomy of the thick leaves in Dendrobium section Rhizobium (Orchidaceae).
Int J Plant Sci.
1994;
155
716-729
-
14
Carlsward B S, Stern W L, Judd W S, Lucansky T W.
Comparative leaf anatomy and systematics in Dendrobium, sections Aporum and Rhizobium (Orchidaceae).
Int J Plant Sci.
1997;
158
332-342
-
15 Anonymous. Chinese material medica (Zhong Yao Zhi). 2nd edition, Vol. 4. Beijing; People's Health Publishing House 1988: 230-446
-
16
Zhang Y B, Wang J, Wang Z T, But P P, Shaw P C.
DNA microarray for identification of the herb of Dendrobium species from Chinese medicinal formulations.
Planta Med.
2003;
69
1172-1174
-
17
Lau D T, Shaw P C, Wang J, But P P.
Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA.
Planta Med.
2001;
67
456-460
-
18
Xu H, Wang Z T, Ding X Y, Zhou K Y, Xu L S.
Differentiation of Dendrobium species used as “Huangcao Shihu” by rDNA ITS sequence analysis.
Planta Med.
2006;
72
89-92
-
19
Lahaye R, van der Bank M, Bogarin D, Warner J, Pupulin F, Gigot G, Maurin O, Duthoit S, Barraclough T G, Savolainen V.
DNA barcoding the floras of biodiversity hotspots.
Proc Natl Acad Sci USA.
2008;
105
2923-2928
-
20
Yip P Y, Chau C F, Mak C Y, Kwan H S.
DNA methods for identification of Chinese medicinal materials.
Chin Med.
2007;
2
9
Prof. Shi-Lin Chen
Institute of Medicinal Plant Development
Chinese Academy of Medical Sciences
No. 151 Malianwa North Road, Haidian District
Beijing 100193
People's Republic of China
Phone: + 86 10 62 89 97 00
Fax: + 86 10 62 89 97 76
Email: slchen@implad.ac.cn
