Phylogenetic Relationships Between Sedum L. and Related Genera (Crassulaceae) Based on ITS Rdna Sequence Comparisons
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Flora 224 (2016) 218–229 Contents lists available at ScienceDirect Flora j ournal homepage: www.elsevier.com/locate/flora Phylogenetic relationships between Sedum L. and related genera (Crassulaceae) based on ITS rDNA sequence comparisons a,∗ a b Vyacheslav Yu. Nikulin , Svetlana B. Gontcharova , Ray Stephenson , a Andrey A. Gontcharov a Institute of Biology and Soil Science FEB RAS, 100-letia Vladivostoka Prosp., 159, Vladivostok, 690022, Russia b Percy Gardens 8, Choppington, Northumberland, NE62 5YH, England, United Kingdom a r t i c l e i n f o a b s t r a c t Article history: Sedum is the most species-rich and taxonomically complex member of the family Crassulaceae. The genus Received 26 May 2016 comprises ca. 420 species, which display notable diversity and homoplasy of growth forms, vegetative Received in revised form 12 August 2016 and reproductive features traditionally used to delineate crassulacean genera therefore it is impossible Accepted 12 August 2016 to characterize Sedum phenotypically. Artificial nature of Sedum was recognized long time ago and it Edited by W. Durka was characterized as a “catch-all” taxon. The phylogenetic structure of Sedum and its relationship with Available online 23 August 2016 allied genera are poorly understood. Two hundred twenty three sequences of nuclear ribosomal internal transcribed spacer (85 were obtained in this study) were used to address this question and provide a Keywords: Crassulaceae phylogeny-based framework for further taxonomic revisions. To identify positional homology between Sedum divergent sequences, secondary structure models were generated and used to guide the alignment. Our ITS rDNA molecular phylogenetic data corroborated results of previous studies based on other markers and datasets Secondary structure and support the monophyly of the tribes Aeonieae, Semperviveae, and Sedeae. Our study is the first to Phylogeny resolve the clade of the tribe Semperviveae based on sequence comparisons, and the clade of the tribe Sedeae with nrDNA. Sedeae accommodated majority of Sedum species either in the robust Acre clade or paraphyletic Leucosedum cluster. Besides Sedum, these lineages comprised a number of genera that were mostly embedded among Sedum species. The highly polyphyletic nature of Sedum and its relatives corroborated in this study using a large taxon set, calls for re-evaluation of the genus concept in the family Crassulaceae and the tribe Sedeae in particular. © 2016 Elsevier GmbH. All rights reserved. 1. Introduction One of the reasons for the poor status of Sedum taxonomy is its extreme morphological diversity and homoplasy of phenotypic The genus Sedum L. is the most species-rich member of the features traditionally used to delineate crassulacean genera. In fact stonecrop family Crassulaceae DC., as well as the morphologically artificial nature of this genus was recognized long before the advent diverse and taxonomically complex. It has significantly reduced in of molecular phylogenetic era, and Sedum was characterized as a the last 50 years, but it still encompasses ca. 420 species that con- “catch-all” taxon (Moran, 1942; Spongberg, 1978; Uhl, 1963). The stitute one third of the family diversity (Thiede and Eggli, 2007). genus concept has changed significantly since then by separating Molecular data unambiguously demonstrate a polyphyletic nature a number of infrageneric taxa into genera, but it is still impossible of the genus with species placed in four major crown clades of to characterize Sedum phenotypically (’t Hart and Bleij, 2005; Mort the crassulacean tree, Acre, Aeonium, Leucosedum, and Sempervivum et al., 2001, 2010; Thiede and Eggli, 2007). Some of these segregates (Gontcharova et al., 2008; van Ham, 1995; van Ham and ’t Hart, (e.g., Graptopetalum Rose, Dudleya Britton et Rose, Prometheum 1998; Mort et al., 2001, 2010). Moreover, at least nine mostly mor- (Berger) Ohba, Sedella Britton et Rose, etc.) show a close relationship phologically distinct crassulacean genera are nested within Sedum. with Sedum species in phylogenetic analyses, thus refuting their independent status or, more plausibly, the current Sedum concept. There is no agreement between specialists regarding the infra- generic structure of Sedum. Up to 30 sections have been recognized in the genus (Alexander, 1942; Berger, 1930; Clausen, 1943; Fu, ∗ Corresponding author. 1965, 1974; ’t Hart, 1991; Jacobsen, 1974; Praeger, 1921; Uhl, 1980), E-mail addresses: [email protected] (V.Yu. Nikulin), of which some have been raised to the subgenus level (Clausen, [email protected] (S.B. Gontcharova), [email protected] 1943, 1979). Fröderström (1930, 1931) proposed seven informal (R. Stephenson), [email protected] (A.A. Gontcharov). http://dx.doi.org/10.1016/j.flora.2016.08.003 0367-2530/© 2016 Elsevier GmbH. All rights reserved. V.Yu. Nikulin et al. / Flora 224 (2016) 218–229 219 groups based on geographic distribution and the type of fruit. At more than one traditional genus. Monophyly was confirmed for least two of these were later described as sections, Filipes (Fröd.) S. only three genera: Lenophyllum, Thompsonella, and Pachyphytum H. Fu and Oreades (Fröd.) K. T. Fu (Fu, 1965, 1974). Two subgenera (Carrillo-Reyes et al., 2009). Thus, Sedum remains poorly studied are currently recognized: Gormania (Britton) Clausen and Sedum (’t phylogenetically. The branching pattern between its clades and lin- Hart and Bleij, 2005; Thiede and Eggli, 2007). Gormania includes 110 eages remains largely unresolved, and only the Acre/Leucosedum species found in the Aeonium, Sempervivum, and Leucosedum clades sistership is undisputable. and mostly distributed in Europe, the Mediterranean region, and ITS rDNA data have been frequently used to access relationships North America (van Ham and ’t Hart, 1998; Mort et al., 2001; Thiede in Crassulaceae at different taxonomic scales, from family-level and Eggli, 2007). The subgenus Sedum accounts for 320 species phylogeny to subsection (Acevedo-Rosas et al., 2004; Carrillo-Reyes comprising the Acre clade and occurring mostly in Asia (ca. 120 et al., 2008, 2009; Gontcharova et al., 2008; Kozyrenko et al., 2013; spp.) and the Americas (ca. 170 spp.; van Ham and ’t Hart, 1998; Mayuzumi and Ohba, 2004; Nikulin et al., 2015), and a great deal Mort et al., 2001). Gormania taxa are usually glandular-pubescent of sequences have been accumulated. However, despite consider- with broadly sessile sepals of equal length and costate seeds, while able interest in the phylogenetic utility of the spacers, relatively Sedum representatives are mostly glabrous with sepals that are free little is known about ITS molecular evolution in Crassulaceae in at the base and spurred (unequal in length when broadly sessile), general and the genus Sedum in particular. High substitution rates as well as reticulate-papillate to reticulate testa. However, there in the spacer may lead to saturation and homoplasy, and fre- are discrepancies in the subgeneric assignment of some taxa (e.g. S. quent length mutations hinder homology assessment. Despite a sedoides (Decaisne) Pau ex Vidal y Lopez, S. hispanicum L., S. gracile relatively rapid rate of primary sequence divergence, conserved C.A. Meyer, etc.; ’t Hart and Bleij, 2005; Thiede and Eggli, 2007). structural elements within the secondary structures of ITS aid the The phylogenetic structure of Sedum and its relationship with accurate alignment of homologous positions (Alvarez and Wendel, allied genera are poorly understood. The genus was generally con- 2003; Coleman, 2003, 2007; Gottschling and Plotner, 2004; Mai sidered as the most primitive and ancient member of the family and Coleman, 1997; Wolf et al., 2005). To clarify the structural evo- (Schönland, 1891), but molecular phylogenetic studies refuted this lution of the ITS region in Sedum and address its phylogeny in a hypothesis by placing its species in a crow assemblage of the more comprehensive way, the ITS regions for 85 accessions were crassulacean tree recognized as the subfamily Sempervivoideae newly sequenced for this study and analyzed together with 138 (Thiede and Eggli, 2007). There, Sedum taxa comprise a bulk of sequences from Genbank. This widely and densely sampled dataset the Acre and Leucosedum clades and are intermixed with mem- facilitated the process of alignment by providing many intermedi- bers of other genera (tribe Sedeae). Several North African species ary sequences. The objectives of this study are 1) to test previously branched paraphyletically at the base of the Aeonium clade (tribe proposed taxonomic and phylogenetic schemes of the crown cras- Aeonieae) comprising Macaronesian genera Aeonium Webb et sulacean assemblage using ITS rDNA sequence variation and 2) to Berth., Aichryson Webb et Berth., and Monanthes Haw. Sedum ser. derive an accurate RNA secondary structure model for this spacer Rupestria Berger (7 species recognized as the genus Petrosedum region for Sedum. Grulich by some students) forms a distinct subclade (or inde- pendent clade) in the Sempervivum clade (tribe Semperviveae; 2. Materials and methods Carrillo-Reyes et al., 2009; Gontcharova et al., 2008; van Ham and ’t Hart, 1998; Mayuzumi and Ohba, 2004; Mort et al., 2001, 2002). 2.1. Plant material Leucosedum was one of the best supported clades in the anal- ysis of chloroplast DNA restriction site data (van Ham, 1995; van The plant material was collected from the field or received Ham and ’t Hart, 1998). However, in all further studies based on from several private collections.