Assessment of relationships between Iranian Fritillaria (Liliaceae) Species Using Chloroplast trnh-psba Sequences and Morphological Characters

Document Type: Research Article


Department of Biology, University of Shahrekord, Shahrekord, Iran


The genus Fritillaria comprises of 165 taxa of medicinal, ornamental and horticultural importance. Evolutionary relationships in this genus is an interesting research area, attracting many researchers. In this study, phylogenetic relationships among 18 native to endemic species in Iran belonging to four subgenera Petilium, Theresia, Rhinopetalum and Fritillaria, are assessed using chloroplast trnH-psbA IGS sequences. Fifteen variable morphological characters are studied, and used in constructing a numerical classification. Results of molecular data showed that subgenus Fritillaria in Iran was a polyphyletic group. Members of the section Olostyleae appeared as paraphyletic. Species non-monophyly was revisited for Fritillaria crassifolia. Both morphological and molecular data show that Fritillaria zagrica and Fritillaria pinardii were closely related taxa, although they may be retain as separate species based on some morphological differences. Multivariate analysis of morphological data arranged the species in consistent groups as with the phylogenetic tree based on sequence data. Results of this study revealed feasibility of the trnH-psbA sequences for contribution in phylogenetic reconstruction in the genus Fritillaria.
Keywords: Fritillaria, Iran, Morphology, Phylogenetic, trnH-psbA


Advay M, Tek┼čen M, Maroofi H. 2015. Fritillaria avromanica sp. nov. (Liliaeceae) from Iran and notes on F. melananthera in Turkey. Nord J Bot first published online.

Aldrich J, Cherney BW, Merlin E, Christopherson L. 1988. The role of insertions/deletions in the evolution of the intergenic region between psbA and trnH in the chloroplast genome. Current Genetics 14: 137-146.

Azuma H, Thien LB, Kawano S. 1999. Molecular phylogeny of Magnolia (Magnoliaceae) inferred from cpDNA sequences and evolutionary divergence of the floral scents. J Plant Res 112: 291-306.

Baker JG. 1874. A revision of the genera and species of Tulipeae. J Linn Soc London, Bot 14: 211-310.

Bakhshi-Khaniki G. 1998. Taxonomy and karyology of the genus Fritillaria s. lat. (Liliaceae) in South West Asia with special reference to the species in Iran. Ph. D. Thesis, Gatteborg University, Sweden.

Bakhshi-Khaniki G. 2002a. Chromosome number of all Iranian species of Fritillaria caucasica group (Liliaceae). Nucleus 45: 103-108.

Bakhshi-Khaniki G. 2002b. Chromosome number of Fritillaria subgenera Petilium and Theresia (Liliaceae). Nucleus 45: 6-11.

Bakhshi-Khaniki G. 2005. Giemsa C-banding studies on interphase nuclei of Iranian species of Fritillaria and Rhinopetalum (Liliaceae). P Natl A Sci India B 75: 294.

Bakhshi-Khaniki G, Persson K. 1997. Nectary morphology in South West Asian Fritillaria (Liliaceae). Nord J Bot 17: 579-611.

Bakhshi-Khaniki G. 1997a. Fritillaria atrolineata (Liliaceae), a new species from Iran. Edinburgh J Bot 54: 171-181.

Bakhshi-Khaniki G. 1997b. Fritillaria chlororhabdota (Liliaceae), a new species from Iran. Herbertia, 52: 140-152.

Bentham G, Hooker JD. 1883. Fritillaria. In: Bentham G., Hooker JD, eds. Genera Plantarum vol. 3. London: Reeve and Co., 817-818.

Boissier E. 1882. Flora Orientalis. vol. 35. H. Georg, Geneva & Basilea, pp. 176-190.

Celebi A, Teksen M, Acik L, Aytac Z. 2008. Taxonomic relationships in genus Fritillaria (Liliaceae): Evidence from RAPD-PCR and SDS-PAGE of seed proteins. Acta Bot Hung 50: 325-343.

Day PD, Berger M, Hill L, Fay MF, Leitch AR, Leitch IJ, Kelly LJ. 2014. Evolutionary relationships in the medicinally important genus Fritillaria L. (Liliaceae). Mol Phylogenet Evol 80: 11-19.

Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemistry 19: 11-15.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39: 783-791.

Fukuda T, Yokoyama J, Tsukaya H. 2003. Phylogenetic relationships among species in the genera Chisocheton and Guarea that have unique indeterminate leaves as inferred from sequences of chloroplast DNA. Int J Plant Sci 164: 13-24.

Funk DJ, Omland KE. 2003. Species-level paraphyly and polyphyly: frequency, causes, and consequences, with insights from animal mitochondrial DNA. Annu Rev Ecol Evol Syst 34: 397-423.

Holdregger R, Abbott RJ. 2003. Phylogeography of the Arctic-Alpine Saxifraga oppositifolia (Saxifragaceae) and some related taxa based on cpDNA and ITS sequence variation. Am J Bot 90: 931–936.

Jafari H, Babaei A, Karimzadeh G, Ahmadi-Roshan M. 2014. Cytogenetic study on some Fritillaria species of Iran. Plant Syst Evol 300: 1373-1383.

Khourang M, Babaei A, Sefidkon F, Naghavi MR, Asgari D, Potter D. 2014. Phylogenetic relationship in Fritillaria spp. of Iran inferred from ribosomal ITS and chloroplast trnL-trnF sequence data. Biochem Syst Ecol 57: 451-457.

Miller JT, Grimes JW, Murphy DJ, Bayer RJ, Ladiges PY. 2003. A phylogenetic analysis of the Acacieae and Ingeae (Mimosoideae: Fabaceae) based on trnK, matK, psbA-trnH, and trnL/trnF sequence data. Syst Bot 28: 558-566.

Ozhatay N. 2000. Fritillaria L. In: Guner A, Ozhatay N, Ekim T, Baser KHC. eds. Flora of Turkey and the East Aegean Islands (suppl. 2). Edinburgh: Edinburgh University Press, 243-246.

Rix EM. 1975. Notes on Fritillaria (Liliaceae) in the Eastern Mediterranean Region, III. Kew Bull 30: 153-162.

Rix EM. 1984. Fritillaria L. In: Davis PH, ed. Flora of Turkey. Edinburgh: 284–302.

Rix EM. 1997. Fritillaria L. (Liliaceae) in Iran. Iran J Bot 1: 75-95.

Rix EM, Frank E, Webster G. 2001. Fritillaria: a revised classification, together with an updated list of species. Fritillaria Group of Alpine Garden Soc., Edinburgh.

Rohlf FJ. 2000. NTSYS-pc: numerical taxonomy and multivariate analysis system, version 2.1. Exeter Software, Setauket, NY.

Ronsted N, Law S, Thornton H, Fay MF, Chase MW. 2005. Molecular phylogenetic evidence for the monophyly of Fritillaria and Lilium (Liliaceae; Liliales) and the infrageneric classification of Fritillaria. Mol Phylogenet Evol 35: 509-527.

Ryan SP. 2014. Molecular phylogeny and character evolution of Fritillaria subgenus Liliorhiza (Liliaceae). Thesis for degree of Master of Science in Biology. San Diego State University. Retrieved from

Rzhetsky A, Nei M. 1992. A simple method for estimating and testing minimum-evolution trees. Mol Biol Evol 9: 945-967.

Sang T, Crawford DJ, Stuessy TF. 1997. Chloroplast DNA phylogeny, reticulate evolution, and biogeography of Paeonia (Paeoniaceae). Am J Bot 84: 1120–1136.

Shaw J, Small RL. 2005. Chloroplast DNA phylogeny and phylogeography of the North American plums (Prunus subgenus Prunus section Prunocerasus, Rosaceae). Am J Bot 92: 2011-2030.

Shaw J, Lickey EB, Beck JT, Farmer SB, Liu W, Miller J, Siripun KC, Winder CT, Schilling EE, Small RL. 2005. The tortoise and the hare II: Relative utility of 21 noncoding chloroplast DNA sequences for phylogenetic analysis. Am J Bot 92: 142-166.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731-2739.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30: 2725-2729.

Teksen M, Aytac Z. 2011. The revision of the genus Fritillaria L. (Liliaceae) in the Mediterranean region (Turkey). Turk J Bot 35: 447-478.

Teksen M, Aytac Z, Pinar NM. 2010. Pollen morphology of the genus Fritillaria L.(Liliaceae) in Turkey. Turk J Bot 34: 397-416.

Townsend CC. 1985. Fritillaria L. In: Townsend C. C. and Guest E. eds. Flora of Iraq. Baghdad: The Whitefriars Press, 42-185.

Turktas M, Aslay M, Kaya E, Ertugrul F. 2012. Molecular characterization of phylogenetic relationships in Fritillaria species inferred from chloroplast trnL-trnF sequences. Turk J Biol 36: 552-560.

Turrill WB, Sealy JR. 1980. Studies in the genus Fritillaria (Liliaceae). Hookers Icones Plantarum 39: 1-2.

Wendelbo P. 1990. Fritillaria. In: Rechinger KH. ed. Flora Iranica. Graz: Akademische Druck-u. Verlagsanstalt, 61-76.

Zarrei M, Wilkin P, Fay MF, Ingrouille MJ, Zarre S, Chase MW. 2009. Molecular systematics of Gagea and Lloydia (Liliaceae; Liliales): implications of analyses of nuclear ribosomal and plastid DNA sequences for infrageneric classification. Ann Bot 104: 125-142.