Morphological and ITS-based Phylogenetic Studies of Lonicera L.(Caprifoliaceae) Species Growing in Iran

Document Type : Research Article


1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran. Iran.

2 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 Research Institute of Forests and Rangelands, Agricultural Research Education and Extension Organization, Tehran, Iran

4 Department of Microbiology and Biology, Parand Branch, Islamic Azad University, Tehran, Iran


Genus Lonicera has members that have opposite, narrowly elliptic to obovate leaves and yellow-white, red, or purple-red corollas together with capitate stigmas and undulated calyxes. According to Wendelbo (1965) in Flora Iranica, the 19 members of Lonicera are classified into two subgenera, i.e., Lonicera and Chamaecerasus, and three sections, i.e., Isika, Isoxylosteum, and Coeloxylosteum. The four studied species belong to subgenus Chamae­cerasus and sections Isika and Coeloxylosteum. The taxonomy and phylogeny of this genus is highly complicated and controversial. The present study was done by the use of phenetic analyses of morphology together with Bayesian analyses of molecular data (ITS sequences) to illustrate the species relationships, taxonomic classification, and monophyly versus paraphyly of the species in genus Lonicera. We used seven Lonicera species for molecular studies, for which nrDNA-ITS sequences were newly obtained. Successive reweighting with rescaled consistency index was used to conduct the molecular examination, which showed close similarities among the results of maximum likelihood, maximum parsimony, and Bayesian methods based on the ITS dataset were observed. This study showed that in general, it is possible to differentiate the species via morphological features. Phylogenetic relationships within Lonicera were revealed, and ITS-based phylogenetic trees and morphological characters were in agreement.


Amini E, Nasrollahi F, Sattarian A, Khormali A, Habibi M. 2019. Micro-morphological and molecular study of four species of Lonicera (Caprifoliaceae) in Iran. Phytol Balc 25(2): 181-190.
Bakker FT, Culham A, Hettiarachi P, Touloumenidou T, Gibby M. 2004. Phylogeny of Pelargonium (Geraniaceae) based on DNA sequences from three genomes. Taxon 53(1): 17-28.
Baldwin BG, Sanderson MJ, Porter JM, Wojciechowski MF, Campbell CS, Donoghue MJ. 1995. The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny. Ann Mo Bot Gard 82: 247-277.
Baldwin BG. 1992. Phylogenetic utility of the internal transcribed spacers of nuclear ribosomal DNA in plants: An example from the Compositae. Mol Phylogenet Evol 1(1): 3-16.
Bouckaert R, Heled J, Kühnert D, Vaughan T, Wu CH, Xie D, Drummond AJ. 2014. BEAST 2: a software platform for Bayesian evolutionary analysis. PLoS Comput Biol 10(4): e1003537.
Bozchaloyi SE, Sheidai M, Keshavarzi M, Noormohammadi Z. 2017a. Genetic diversity and morphological variability in Geranium purpureum vill. (Geraniaceae) Of Iran. Genetika 49(2): 543-557.
Bozchaloyi SE, Sheidai M, Keshavarzi M, Noormohammadi Z. 2017b. Species delimitation in Geranium Sect. Batrachioidea: Morphological and Molecular Acta Bot Hung 59(3-4): 319-334.
Bozchaloyi SE, Sheidai M, Keshavarzi M, Noormohammadi Z. 2017c. Genetic and morphological diversity in Geranium dissectum (Sec. Dissecta, Geraniaceae) populations. Biologia 72(10): 1121-1130.
Bozchaloyi SE, Sheidai M, Keshavarzi M, Noormohammadi Z. 2017d. Analysis of genetic diversity in Geranium robertianum by ISSR markers Phytol Balc 23(2): 157-166.
Bozchaloyi SE, Sheidai M, Keshavarzi M. Noormohammadi Z. 2018. Species relationship and population structure analysis in Geranium subg. Robertium with the use of ISSR molecular markers. Acta Bot Hung 60(1-2):47-65
Chase MW, Soltis DE, Olmstead RG, Morgan D, Les DH, Mishler BD, Albert VA. 1993. Phylogenetics of seed plants: an analysis of nucleotide sequences from the plastid gene rbcL. Ann Missouri Bot Gard 80(3): 528-580.
Cunningham CW. 1997. Can three incongruence tests predict when data should be combined? Mol Biol Evol 14:733-740
Farris JS, Kallersjo M, Kluge AG, Bult C. 1995. Testing significance of incongruence. Cladistics 10:315-319.
Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39(4):783-791
Foulds LR, Robinson DF. 1981. Comparison of phylogenetic trees. Math Biosci 53(1-2): 131-147.
Ghahremaninejad F, Ezazi A. 2009. A new record for the flora of Iran: Lonicera microphylla (Caprifoliaceae). Iran J Bot 15(2): 157-159.
Hammer Ø, Harper D, Ryan PD. 2012. PAST: Paleontological Statistics software package for education and data analysis. Palaeontol Electron 4(1): 1-9.
Hara H. 1983. A revision of Caprifoliaceae of Japan with reference to allied plants in other districts and the Adoxaceae. Ginkgoana, No. 5, Contributions to the flora of Asia and the Pacific region. Tokyo: Academia Scientific Books.
Hsu PS, Wang HJ. 1988. Lonicera Linn. In: Flora Republicae Popularis Sinicae. Beijing 72: 143-259.
Judd WS, Campbell CS, Kellogg EA, Stevens PF, Donoghue MJ. 2008. Plant systematics: A phylogenetic approach, 3rd eds. Sunderland, MA: Sinauer.
Li Y, Cai W, Weng X, Li Q, Wang Y, Chen Y, Wang H. 2015. Lonicerae Japonicae Flos and Lonicerae Flos: a systematic pharmacology review. Evid Based Complement Alternat Med 2015: 905063. doi: 10.1155/2015/905063.
Mabberley DJ. 2008. The Plant Book, a Portable Dictionary of Higher Plants. Cambridge Univ Press, Cambridge.
Nakaji M, Tanaka N, Sugawara T. 2015. A molecular phylogenetic study of Lonicera L. (Caprifoliaceae) in Japan based on chloroplast DNA sequences. Acta Phytotaxon Geobot 66(3): 137-151.
Naugžemys D, Žilinskaitė S, Kleizaitė V, Skridaila A, Žvingila D. 2011. Assessment of genetic variation among elite and wild germplasm of blue honeysuckle (Lonicera caerulea L). Balt For 17: 8-16.
Nuin P. 2005. MrMTgui 1.0 (version 1.6). Program distributed by the author at
Nylander JAA. 2004. MrModeltest v2. Program distributed by the author. Evolutionary Biology Centre, Uppsala University
Osaloo SK, Maassoumi AA, Murakami N. 2003. Molecular systematics of the genus Astragalus L. (Fabaceae): Phylogenetic analyses of nuclear ribosomal DNA internal tran­scribed spacers and chloroplast gene ndhF sequences. Plant Syst Evol 242(1): 1-32.
Osaloo SK, Maassoumi AA, Murakami N. 2005. Molecular systematics of the Old World Astragalus (Fabaceae) as inferred from nrDNA ITS sequence data. Brittonia 57(4): 367-381.
Podani J. 2000. Introduction to the Exploration of Multivariate Data [English translation]. Leide, Netherlands: Backhuyes.
Posada D, Backley TR. 2004. Model selection and model averaging in phylogenetics: advantages of akaike information criterion and Bayesian approaches over likelihood ratio tests. Syst Biol 53(5):793-808.
Ronquist F, Huelsenbeck JP (2003) MrBayes 3: bayesian phylogenetic inference under mixed models. Bioinformatics 19(12):1572-1574.
Shang X, Pan H, Li M, Miao X, Ding H. 2011. Lonicera japonica Thunb: Ethnopharmacology, phytochemistry and pharmacology of an important traditional Chinese medicine. J Ethnopharmacol 138(1): 1-21.
Smolik M, Ochmian I, Grajkowski J. 2010. Genetic variability of Polish and Russian accessions of cultivated blue honeysuckle (Lonicera caerulea). Russ J Genet 46(8): 960-966.
Soltis ED, Soltis P. 2000. Contributions of plant molecular systematics to studies of molecular evolution. Plant Mol Biol 42(1): 45-75.
Swofford DL. 2002. PAUP: phylogenetic analysis using parsimony (and other methods), Version 4.0, Beta 10. Sinauer Associates, Sunderland.
Taberlet P, Gielly L, Pautou G, Bouvet J. 1991. Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17(5):1105-1109.
Theis N, Donoghue JM, Li J. 2008. Phylogenetics of the Caprifolieae and Lonicera (Dipsacales) based on nuclear and chloroplast DNA sequences. Syst Bot 33(4): 776-783.
Wendelbo P. 1965. Caprifoliaceae in Rechinger, KH (ed.): Flora Iranica, no. 10. Graz: Akademische Druck-und Verlagsanstalt, 159.
White TJ, Bruns T, Lee SJWT, Taylor JW. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: a guide to methods and applications 18(1): 315-322.
Zeng H, Li Y, Chen J, Wang X, Qian Z, Li Y, Cai N, Liu S. 2017. Lonicera japonica ‘Fenglei’. HortScience 52(5): 789-791.
Zhang Q, Feild TS, Antonelli A. 2015. Assessing the impact of phylogenetic incongruence on taxonomy, floral evolution, biogeographical history, and phylogenetic diversity. Am J Bot 102(4): 566-580.