Genetic Diversity in Hypericum and AFLP Markers for Species-Specific Identification of H. perforatum L.

Abstract

One of the top-selling medicinal products worldwide is Hypericum perforatum (St. John’s Wort). Despite its cosmopolitan distribution and utilization, little is known regarding the relationship of the bioactive compounds in H. perforatum to the plants from which they are purportedly derived. In this study, amplified fragment length polymorphism (AFLP) analysis of 56 Hypericum accessions, representing 11 species, was conducted to gain a better understanding of diversity within Hypericum species, especially within cultivated accessions of H. perforatum, and to establish a molecular methodology that will provide breeders and regulators with a simple, affordable, and accurate tool with which to identify purported H. perforatum material. Utilizing four primer combinations, a total of 298 polymorphic markers were generated, of which 17 were present in all H. perforatum accessions and 2 were specific to only H. perforatum. This study demonstrates that AFLP can be utilized not only to determine the relationships of closely related Hypericum accessions, but as a tool to authenticate material in herbal remedies through the use of genetic fingerprinting.

Key words

Hypericum - Hypericaceae - marker analysis - genetic diversity - St. John’s wort

References

• 1 Robson N. Hypericum botany. In: Ernst E, editor The genus Hypericum. London; Taylor and Francis 2003: 1-22.
• 2 Robson N KB. Studies in the genus Hypericum L. (Guttiferae) 1. Infrageneric classification. Bull Brit Mus (Natural History, Botany series) 1977: 291-355.
• 3 Campbell M H, Delfosse E S. The biology of Australian weeds. 13. Hypericum perforatum L.  J Aust I Agr Sci. 1984;  50 63-73.
• 4 Martonfi P, Brutovska R, Cellarova E, Repcak M. Apomixis and hybridity in Hypericum perforatum .  Folia Geobot Phytotaxon. 1996;  31 389-96.
• 5 Matzk F, Meister A, Brutovska R, Schubert I. Reconstruction of reproductive diversity in Hypericum perforatum L. opens novel strategies to manage apomixis.  Plant J. 2001;  26 275-82.
• 6 Barcaccia G, Arzenton F, Sharbel T F, Varotto S, Parrini P, Lucchin M. Genetic diversity and reproductive biology in ecotypes of the facultative apomict Hypericum perforatum L.  Heredity. 2006;  96 322-34.
• 7 Mayo G M. Modes of reproduction in Australian populations of Hypericum perforatum L. (St. John’s wort) revealed by DNA fingerprinting and cytological methods.  Genome. 2003;  46 573-9.
• 8 Axarlis S, Mentis A, Demetzos C, Mitaku S, Skaltsounis A L, Marselos M. et al . Antiviral in vitro activity of Hypericum perforatum L. extract on the human cytomegalovirus (HCMV).  Phytother Res. 1998;  12 507-11.
• 9 Barnes J, Anderson L A, Phillipson J D. St John’s wort (Hypericum perforatum L.): a review of its chemistry, pharmacology and clinical properties.  J Pharm Pharmacol. 2001;  53 583-600.
• 10 Greeson J M, Sanford B, Monti D A. St. John’s wort (Hypericum perforatum): a review of the current pharmacological, toxicological, and clinical literature.  Psychopharmacology. 2001;  153 402-14.
• 11 Nahrstedt A, Butterweck V. Biologically active and other chemical constituents of the herb of Hypericum perforatum L.  Pharmacopsychiatry. 1997;  30 129-34.
• 12 Campbell M H, May C E, Southwell I A, Tomlinson J D, Michael P W. Variation and varietal determination in Hypericum perforatum L. (St. John’s wort) in New South Wales.  Plant Prot Q. 1992;  7 43-5.
• 13 Southwell I A, Campbell M H. Hypericin content variation in Hypericum perforatum in Australia.  Phytochemistry. 1991;  30 475-8.
• 14 Aziz N, Sauve R J, Long D, Cherry M. Genetic and phytochemical diversity assessment among eleven Hypericum accessions via AFLP and HPLC analyses.  J Herbs Spices Med Plants.. 2006;  12 97-105.
• 15 Arnholdt-Schmitt B. RAPD analysis: a method to investigate aspects of the reproductive biology of Hypericum perforatum L.  Theor Appl Genet. 2000;  100 906-11.
• 16 Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M. et al . AFLP: a new technique for DNA fingerprinting.  Nucleic Acids Res. 1995;  23 4407-14
• 17 Gaudeul M, Taberlet P, Till-Bottraud I. Genetic diversity in an endangered alpine plant, Eryngium alpinum L. (Apiaceae), inferred from amplified fragment length polymorphism markers.  Mol Ecol. 2000;  9 1625-37.
• 18 Soleimani V D, Baum B R, Johnson D A. AFLP and pedigree-based genetic diversity estimates in modern cultivars of durum wheat [Triticum turgidum L. subsp. durum (Desf.) Husn.]  Theor Appl Genet. 2002;  104 350-7.
• 19 Blears M J, Grandis S A, Lee H, Trevors J T. Amplified fragment length polymorphism (AFLP): a review of the procedure and its applications.  J Ind Microbiol Biotechnol. 1998;  21 99-114.
• 20 Gustafsson M HG, Bittrich V, Stevens P F. Phylogeny of Clusiaceae based on rbcL sequences.  Int J Plant Sci. 2002;  163 1045-54.
• 21 Robson N. Studies in the genus Hypericum L. (Guttiferae) 4(2). Section 9. Hypericum sensu lato (part 2): subsection 1. Hypericum series 1. Hypericum .  Bull Brit Mus (Natural History, Botany series). 2002;  32 61-123.
• 22 Hawkins J S, Pleasants J, Wendel J F. Identification of AFLP markers that discriminate between cultivated cotton and the Hawaiian island endemic, Gossypium tomentosum Nuttall ex Seeman.  Genet Resour Crop Evol. 2005;  52 1069-78.
• 23 Benham J, Jeung J U, Jasieniuk M, Kanazin V, Blake T. Genographer: a graphical tool for automated fluorescent AFLP and microsatellite analysis. J Agric Genomics 1999: 1-3.
• 24 Swafford D L. Phylogenetic analysis using parsimony (*and other methods), version 4. Sunderland; Sinauer 2002.
• 25 Nei M. Genetic distance between populations.  Am Nat. 1972;  106 283-92.
• 26 Rohlf F J. NTSYS-pc. Numerical taxonomy and multivariate analysis system, Version 2.10. New York; Exeter Software 2002.
• 27 Excoffier L, Smouse P E, Quattro J M. Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data.  Genetics. 1992;  131 479-91.
• 28 Peakall R, Smouse P E. GENEALEX 6: genetic analysis in Excel. Population genetic software for teaching and research.  Mol Ecol Notes. 2006;  6 288-95.
• 29 Maron J L, Vila M, Bommarco R, Elmendorf S, Beardsley P. Rapid evolution of an invasive plant.  Ecol Monogr. 2004;  74 261-80.
• 30 Crockett S L, Douglas A W, Scheffler B E, Khan I A. Genetic profiling of Hypericum (St. John’s wort) species by nuclear ribosomal ITS sequence analysis.  Planta Med. 2004;  70 929-35.