The Phytochemical and Genetic Survey of Common and Dwarf Juniper (Juniperus communis and Juniperus nana) Identifies Chemical Races and Close Taxonomic Identity of the Species

Abstract

Juniperus communis L. (= J. communis var. communis) and Juniperus nana Willd. (= J. communis var. saxatilis) are subspecies of juniper. J. communis grows widely in both hemispheres, primarily in lower elevations while J. nana is mainly observed in high mountains. Although they can be distinguished by morphological features, it is not known whether they are genetically and phytochemically distinct entities. We aimed to check whether it is possible to distinguish these two plants (i) by pharmaceutically important chemical traits and (ii) on the basis of intraspecifically highly polymorphic fragment of chloroplast DNA. We used GC with achiral as well as with enantioselective stationary phase columns to identify the main monoterpenes of the essential oil. Sequence analysis of the trnL (UAA)-trnF (GAA) intergenic spacer of the chloroplast genome was used as a genetic marker of taxonomic identity between these two subspecies. The chromatographic analysis showed the existence of three chemical races - the α-pinene type, the sabinene type and one with intermediate contents of these terpenes among both J. communis and J. nana. Surprisingly, sequence analysis of trnL (UAA)-trnF (GAA) revealed 100 % similarity between the common and the dwarf juniper. Thus, the monoterpene pattern is related to geographical origin, and not to the species identity. We suggest that the three chemical races identified in the present study should be considered as separate sources of pharmaceutical raw material. Our results demonstrate that the contents of α-pinene and sabinene may be applied as a quick diagnostic test for preeliminary evaluation of plant material.

References

• 1 Tutin T G, Heywood V H, Burges N A, Valentine D H, Walters S M, Webb D A. Flora Europea. Cambridge; University Press 1964: p 38-9
• 2 Farjon A. World checklists and bibliography of conifers. Kew; Royal Botanic Gardens 2001: p 298
• 3 Chatzopoulou P S, Katsiotis S T. Procedures influencing the yield and the quality of the essential oil from Juniperus communis berries.  Pharm Acta Helv. 1995;  70 247-53
• 4 Hiltunen R, Laakso I. Gas chromatographic analysis and biogenetic relationships of monoterpene enantiomers in Scots pine and juniper needle oils.  Flavour Fragr J. 1995;  10 203-10
• 5 Proenca da Cunha A, Roque O LR. The chemical composition of the essential oil and alcoholic extract of Juniperus communis L. ssp. nana Syme.  J Ess Oil Res. 1989;  1 15-6
• 6 Looman A, Svendsen A B. The needle essential oil of Norwegian mountain juniper, Juniperus communis L. var. saxatilis Pall.  Flavour Fragr J. 1992;  7 23-6
• 7 Adams R P, Demeke T. Systematic relationships in Juniperus based on random amplified polymorphic DNAs (RAPDs).  Taxon. 1993;  42 553-71
• 8 Adams R P, Pandey R N, Leverenz J W, Dignard N, Hoegh K, Thorfinnsson T. Pan-arctic variation in Juniperus communis: historical biogeography based on DNA fingerprinting.  Biochem Syst Ecol. 2003;  31 181-92
• 9 Zhu S, Fushimi H, Cai S Q, Komatsu K. Phylogenetic relationship in the Genus Panax: inferred from chloroplast trnK gene and nuclear 18S rRNA gene sequences.  Planta Med. 2003;  69 647-53
• 10 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
• 11 Terry R G, Nowak R S, Tausch R J. Genetic variation in chloroplast and nuclear ribosomal DNA in Utah juniper (Juniperus osteosperma, Cupressaceae): evidence for interspecific gene flow.  Am J Bot. 2000;  87 250-8
• 12 Kojoma M, Kurihara K, Yamada K, Sekita S, Satake M, Iida O. Genetic identification of cinnamon (Cinnamomum spp.) based on the trnL-trnF chloroplast DNA.  Planta Med. 2002;  68 94-6
• 13 Yang M, Zhang D, Liu J, Zheng J. A molecular marker that is specific to medicinal rhubarb based on chloroplast trnL/trnF sequences.  Planta Med. 2001;  67 84-6
• 14 Long C, Kakiuchi N, Takahashi A, Komatsu K, Cai S, Mikage M. Phylogenetic analysis of the DNA sequence of the non-coding region of nuclear ribosomal DNA and chloroplast of Ephedra plants in China.  Planta Med. 2004;  70 1080-4
• 15 Vernin G, Boniface C, Metzger J, Ghiglione C, Hammoud A, Suon K N. et al . GC-MS-specma bank analysis of Juniperus communis needles and berries.  Phytochemistry. 1988;  27 1061-4
• 16 Ochocka J R, Asztemborska M, Zook D R, Sybilska D, Perez G, Ossicini L. Enantiomers of monoterpenic hydrocarbons in essential oils from Juniperus communis .  Phytochemistry. 1997;  44 869-73
• 17 Gordon D, Abajian C, Green P. Consed: a graphical tool for sequence finishing.  Genome Res. 1998;  8 195-202
• 18 Ewing B, Hillier L, Wendl M C, Green P. Base-calling of automated sequencer traces using Phred. I. Accuracy assessment.  Genome Res. 1998;  8 175-85
• 19 Hall T A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT.  Nucleic Acids Symp Ser. 1999;  41 95-8