Abstract
Twenty-four Ephedra plants belonging to 8 species grown in the northern and western parts of China were phylogenetically analyzed for their non-coding DNA sequences, internal transcribed spacers (ITSs) of nuclear ribosomal DNA as well as trnL intron and intergenic spacers between trnL and trnF (trnL/trnF) of the chloroplast. Based on the ITS sequences, the 8 species could be divided into 3 groups: Group 1 (Ephedra intermedia, E. sinica, E. przewalskii), Group 2 (E. equisetina, E. monosperma, E. gerardiana), and Group 3 (E. likiangensis, E. minuta). The species classified into Group 1 grow mainly in the north, Group 3 in the south and Group 2 in the center, suggesting their genetic and geographic relationships. A specific primer set was designed to classify the 3 groups by routine PCR. Combined analysis of ITS and trnL/trnF differentiated the 8 Ephedra species.
Abbreviations
ITS1:internal transcribed spacer 1
ITS2:internal transcribed spacer 2
trnL/trnF:intron of trnL and intergenic spacer between trnL and trnF
EI:Ephedra intermedia
ES:Ephedra sinica
EP:Ephedra przewalskii
EE:Ephedra equisetina
EM:Ephedra monosperma
EG:Ephedra gerardiana
EL:Ephedra likiangensis
EMu:Ephedra minuta
Key words
Ephedra
- Ephedraceae - nuclear ribosomal DNA - internal transcribed spacer (ITS) - chloroplast DNA - intron of trnL and intergenic spacer between trnL and trnF (trnL/trnF)
References
-
1 Chinese Pharmacopoeia Editorial C ommittee. Chinese Pharmacopoeia. 2000 edition, Volume 1 Beijing; Chemical Technology Press 2000: p 262
-
2 Zhonghua Bencao (Chinese Herb) Editorial C ommittee. Zhonghua Bencao (Chinese Herb). Volume 2 Shanghai; Shanghai Science & Technology Press 1999: pp 349-357
-
3
Zhang J S, Li S H, Lou Z C.
Morphological and histological studies of Chinese Ephedra mahuang I. Seven species produced in north China.
Acta Pharmaceutica Sinica.
1989;
24
937-48
-
4
Zhang J S, Li S H, Lou Z C.
Morphological and histological studies of Chinese Ephedra mahuang II. Species produced in southwestern China and other species.
Acta Pharmaceutica Sinica.
1990;
25
54-65
-
5
Mikage M, Kondo N.
Medico-botanical studies of Ephedra plants from the Himalayan region, part I. Anatomical studies of the herbal stems and botanical origin of Tibetan crude drugs ”TSHE” and ”BALU”.
J Jpn Bot.
1996;
71
323-32
-
6
Takeuchi M, Nakashima A, Mizukami H, Hiraoka N, Kohda H.
RAPD analysis of Ephedra plants.
Natural Medicines.
2003;
57
50-4
-
7
Mikage M, Takahashi A, Chen H B, Li Q S.
Studies of Ephedra plants in Asia. Part 1. On the resources of Ephedra plants in China.
Natural Medicines.
2003;
57
202-8
-
8
Lau D TW, Shaw P C, Wang J, But P PH.
Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA.
Planta Medica.
2001;
67
456-60
-
9
Ding X Y, Xu L S, Xu H, Wang Z T, Zhou K Y.
Morphological and DNA molecular evidence for authentication of Dendrobium flexicaule from ITS allied species of Dendrobium
.
Acta Pharmaceutica Sinica.
2001;
36
868-73
-
10
Chen Y Q, Wang N, Zhou H, Qu L H.
Differentiation of medicinal Cordyceps species by rDNA ITS sequence analysis.
Planta Medica.
2002;
68
635-9
-
11
Wen J, Zimmer E A.
Phylogeny and biogeography of Panax L. (the Ginseng genus, Araliaceae): inferences from ITS sequences of nuclear ribosomal DNA.
Mol Phylogenet Evol.
1996;
6
167-77
-
12
Liston A, Robinson W A, Oliphant J M.
Length variation in the nuclear ribosomal DNA internal transcribed spacer region of non-flowering seed plants.
Syst Botany.
1996;
21
109-20
-
13
Maggini F, Marrocco R, Gelati M T, Dominicis R ID.
Lengths and nucleotide sequences of the internal spacers of nuclear ribosomal DNA in gymnosperms and pteridophytes.
Pl Syst Evol.
1998;
213
199-205
-
14
Yang M H, Zhang D M, Liu J Q, Zheng J H.
A molecular marker that is specific to medicinal rhubarb based on chloroplast trnL/trnF sequences.
Planta Medica.
2001;
67
784-6
-
15
Mogensen H L.
The hows and whys of cytoplasmic inheritance in seed plants.
Am J Bot.
1996;
83
383-404
-
16
Zhu S, Fushimi H, Cai S Q, Komatsu K.
Phylogenetic relationship in the Genus Panax: inferred from choroplast trnK gene and nuclear 18S rRNA gene sequences.
Planta Medica.
2003;
69
647-653
-
17
Taberlet P, Gielly L, Pautou G, Bouvet J.
Universal primers for amplification of three non-coding regions of chloroplast DNA.
Plant Mol Biol.
1991;
17
1105-9
-
18
Price R A.
Systematics of the Gnetales: a review of morphological and molecular evidence.
Int J Plant Sci.
1996;
157
40-9
-
19 Agenda academiae sinicae e dita. Flora Reipublicae Popularis Sinicae. Volume 7 Beijing; Science Press 1978: pp 468-89
-
20
Chouhdry A S.
Karyomorphological and cytogenetical studies in Ephedra
.
J Sci Hiroshima Univ Ser B, Div 2.
1984;
19
57-109
Dr. N. Kakiuchi
Faculty of Pharmaceutical Sciences
Kanazawa University
13-1 Takaramachi
Kanazawa 920-0934
Japan
Fax: +81-76-234-4490
Email: kakiuchi@p.kanazawa-u.ac.jp