Systematics of Water Lilies (Genus Nymphaea L.) Using 18S rDNA Sequences
Water lily (Nymphaea L.) is the largest genus of Nymphaeaceae. This family is composed of six genera (Nuphar, Ondinea, Euryale, Victoria, Barclaya, Nymphaea). Its members are nearly worldwide in tropical and temperate regions. The classification of some species in Nymphaea is ambiguous due to high variation in leaf and flower parts such as leaf margin, stamen appendage. Therefore, the phylogenetic relationships based on 18S rDNA were constructed to delimit this genus. DNAs of 52 specimens belonging to water lily family were extracted using modified conventional method containing cetyltrimethyl ammonium bromide (CTAB). The results showed that the amplified fragment is about 1600 base pairs in size. After analysis, the aligned sequences presented 9.36% for variable characters comprising 2.66% of parsimonious informative sites and 6.70% of singleton sites. Moreover, there are 6 regions of 1-2 base(s) for insertion/deletion. The phylogenetic trees based on maximum parsimony and maximum likelihood with high bootstrap support indicated that genus Nymphaea was a paraphyletic group because of Ondinea, Victoria and Euryale disruption. Within genus Nymphaea, subgenus Nymphaea is a basal lineage group which cooperated with Euryale and Victoria. The other four subgenera, namely Lotos, Hydrocallis, Brachyceras and Anecphya were included the same large clade which Ondinea was placed within Anecphya clade due to geographical sharing.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.2643860Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 287
 C. Linnaeus, “Species plantarum, 2 vols.” Stockholm, 1753.
 P. P. Gupta, “Cytogenetics of aquatic ornamentals. VI. Evolutionary trends and relationships in the genus Nymphaea,” Cytologia, 45, 1980, pp. 307-314.
 T. Borsch, K. Hilu, J. Wiersema, C. Lohne, W. Barthlott, and V. Wilde, “Phylogeny of Nymphaea (Nymphaeaceae): evidence from substitutions and microstructural changes in the chloroplast trnT-trnF region,” Int. J. Plant Sci., 168(5), 2007. pp. 639–671.
 P. D. Slocum, “Waterlilies and Lotuses. Portland: Cambridge,” 2005.
 Z. K. Shinwari and S. Shinwari, “Molecular data and phylogeny of family Smilacaceae,” Pak. J. Bot., Special Issue (S.I. Ali Festschrift) 42, 2010, pp. 111-116.
 N. C. Andreasen, “Evaluation of brain imaging techniques in mental illness,” Annu. Rev. Med., 39, 1988, pp. 335-345.
 M. Moudi and R. Go, “The comparison between nuclear ribosomal DNA and chloroplast DNA in molecular systematic study of four sections of genus Dendrobium sw. (Orchidaceae),” Int. J. Bioassays, 5(3), 2016, pp. 4944-4952.
 M. A. Buchheim, and R. L. Chapman, “Phylogeny of the colonial green flagellates: a study of 18S and 26S rRNA sequence data,” BioSystems, 25, 1991, pp. 85–100.
 M. A. Buchheim, and R. L. Chapman, “Phylogeny of the colonial green flagellates: A study of 18S and 26S rRNA sequence data,” BioSystems. 25, 1991, pp. 85-100.
 R.L. Chapman, M.A. Buchheim, C.F. Delwiche, T. Friedl, V.A.R. Huss, K.G. Karol, L.A. Lewis, J. Manhart, R.M. McCourt, J.L. Olsen, and D.A. Waters, “Molecular systematics of the green algae. In: The molecular systematics of plants II,” (Eds.): P.S. Soltis, D.E. Soltis, and J.J. Doyle, Kluwer Academic Publishers, Norwell, Massachusetts, USA, 1998, pp. 508-540.
 T.A. Hedderson, R.L. Chapman, and W.L. Rootes, “Phylogenetic relationships of bryophytes inferred fron small-subunit rRNA gene sequences,” Plant Syst. Evol., 200, 1996, pp. 213-224.
 S.M. Chaw, H. Long, B.S. Wang, A. Zharkikh, and W.H. Li, “The phylogenetic position of Taxaceae based on 18S rRNA sequences,” J. Mol. Evol., 37, 1993, pp. 624-630.
 S.M. Chaw, H.M. Sung, H. Long, A. Zharkikh, and W.H. Li, “The phylogenetic positions of the conifer genera Amentotaxus, Phyllocladus, and Nageia inferred from 18S rRNA sequences,” J. Mol. Evol., 41, 1995, pp. 224-230.
 S.M. Chaw, A. Zharkikh, H.M. Sung, T.C. Lau, and W.H. Li, “Molecular phylogeny of gymnosperms and seed plant evolution: analysis of 18S rRNA sequences,” Mol. Biol. Evol., 14, 1997, pp. 56–68.
 D. E. Soltis, and P. S. Soltis, “Phylogenetic relationships among Saxifragaceae sensu lato: a comparison of topologies based on 18S rDNA and rbcL sequences,” Am. J. Bot., 84, 1997, pp. 504–522.
 J. Doyle, and J. Doyle, “A rapid DNA isolation procedure for small quantities of fresh leaf tissue,” Phytochemical Bulletin, 19, 1987, pp. 11-15.
 G.K. Agrawal, R.N. Pandey, and V.P. Agrawal, “Isolate of DNA from Cheorospondias asillaris leaves,” Biotechnol. Biodiversity Lett., 2, 1992, pp. 19-24.
 C. McCarthy, “Chromas version 1.45.” School of Health science, Griffifth University, Gold Coast Campus, Queensland, Australia, 1996.
 K.B. Nicholas, and H.J.B. Nicholas, “GeneDoc: a tool for editing and annotating multiple sequence alignments,” Retrieved November 2, 2011, from http://en.biosoft.net/. 1997.
 J.D. Thompson, T.J. Gibson, F. Plewniak, F. Jeanmougin, D.G. Higgins, “The CLUSTAL_X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools,” Nucleic Acids Res., 25:4876e4882, 1997.
 K. Tamura, D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar, “MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods,” Mol. Biol. Evol., 28, 2011, pp. 2731-2739.
 H.S. Conard, “The Waterlilies: A Monograph of the Genus Nymphaea. Washington: Carnegie Institution,” 1905.
 P.D. Slocum, “Waterlilies and Lotuses.” Portland, Cambridge.
 D.H. Les, E.L. Schneider, D.J. Padgett, P.S. Soltis, D.E. Soltis, and M. Zanis, “Phylogeny, Classification and Floral Evolution of Water Lilies (Nymphaeaceae; Nymphaeales): A synthesis of Non-molecular, rbcL, matK and 18S rDNA Data,” Syst. Bot., 24(1), 1999, pp. 28-46.
 H.A. Begum, K.K. Ghosal, and T.K. Chattopadhyay, “Comparative morphology and floral biology of three species of the genus of Nymphaea from Bangladesh,” Bangl. J. Bot., 39(2), 2010, pp. 179-183.
 C. Lohne, T. Borsch, S.W.L. Jacobs, C.B. Hellquist, and J.H. Wiersema, “Nuclear and plastid DNA sequences reveal complex reticulate patterns in Australian water-lilies (Nymphaea subgenus Anecphya, Nymphaeaceae),” Aust. Systematic. Botany., 21, 2008, pp. 229-250.
 T. Borsch, C. Lohne, M.S. Mbaye, and J. Wiersema, “Towards a complete species tree of Nymphaea: shedding further light on subg. Brachyceras and its relationships to the Australian waterlilies. Telopea,” 13(1-2), 2011, pp. 193-217.
 T. Yamada, R. Imaichi, and M. Kato, “Developmental morphology of ovules and seeds of Nymphaeales,” Am. J. Bot., 88(6), 2001, pp. 963-974.
 M.E. Collinson, “Recent and Tertiary Seeds of the Nymphaeaceae sensu lato with a Revision of Brasenia ovula (Brong.) Reid and Chandler,” Ann. Bot., 46, 1980, pp. 603-632.
 E.M. Friis, K.R. Pedersen, and P.R. Crane, “Early angiosperm diversification: the diversity of pollen associated with angiosperm reproductive structures in Early Cretaceous floras from Portugal,” Ann. Missouri Bot. Gard., 86, 1999, pp. 259-296.
 M. Ito, “Phylogenetic Systematics of the Nymphaeales,” The botanical magazine, 100, 1987, pp. 17-35.