A multi-locus molecular phylogeny of the Lepidoziaceae: Laying the foundations for a stable classification Endymion D. Cooper a,b,⇑ , A. Jonathan Shaw c , Blanka Shaw c , Murray J. Henwood a , Margaret M. Heslewood b , Elizabeth A. Brown b a School of Biological Sciences, University of Sydney, NSW 2006, Australia b National Herbarium of New South Wales, Mrs Macquaries Road, Sydney NSW 2000, Australia c Department of Biology, Duke University, Durham, NC 27708, USA article info Article history: Received 10 November 2010 Revised 31 January 2011 Accepted 2 February 2011 Available online 18 February 2011 Keywords: Lepidoziaceae Jungermanniopsida Molecular phylogeny Classification Neogrollea Zoopsidioideae abstract The Lepidoziaceae, with over 700 species in 30 genera, is one of the largest leafy liverwort families. Despite receiving considerable attention, the composition of subfamilies and genera remains unsatisfac- torily resolved. In this study, 10 loci (one nuclear 26S, two mitochondrial nad1 and rps3, and seven chlo- roplast atpB, psbA, psbT-psbH, rbcL, rps4, trnG and trnL-trnF) are used to estimate the phylogeny of 93 species of Lepidoziaceae. These molecular data provide strong evidence against the monophyly of three subfamilies; Lepidozioideae, Lembidioideae and Zoopsidoideae, and seven of the 20 sampled genera; Lepidozia, Telaranea, Kurzia, Zoopsis, Lembidium, Paracromastigum and Chloranthelia. Several robust clades are recognised that might provide the basis for a revised subfamily circumscription including a narrower circumscription of the Lepidozioideae and a more inclusive Lembidioideae. Neogrollea notabilis is returned to the Lepidoziaceae and Megalembidium insulanum is placed in the Lembidioideae. Ó 2011 Elsevier Inc. All rights reserved. 1. Introduction The Lepidoziaceae is one of the largest families of leafy liver- worts (Jungermanniales) with over 2200 published binomials, but perhaps as few as 720 accepted species, in 30 genera (Crandall- Stotler et al., 2009; Müller, 2007). With a cosmopolitan distribution, the family has its greatest species diversity in the southern hemi- sphere leading some to postulate a Gondwanan origin (Schuster, 2000), which is consistent with an early to mid-cretaceous origin of the family (Heinrichs et al., 2007). Unrivalled in the breadth of gametophyte morphological diversity, the Lepidoziaceae are united by conserved sporophyte morphology and isophyllous gynoecial branches (Schuster, 2000). Nevertheless, the morphological con- trast between the robust leafy shoots of Bazzania and Lepidozia and the lax-celled, pseudo-thalli of Zoopsis and Zoopsidella, was suf- ficient for some authors to divide the taxa amongst six families (Evans, 1939; Fulford, 1963a, 1966, 1968; Nakai, 1943). Further- more, the degree of gametophyte polymorphism within genera of the Lepidoziaceae is at times greater than that exhibited by entire families of Jungermanniales (Schuster, 2000). This morphological diversity is accompanied by an equally broad range of ecological differentiation, with plants occurring in habitats ranging from san- dy alpine soils to lowland tree trunks. Recent treatments of the Marchantiophyta (Crandall-Stotler and Stotler, 2000; Crandall-Stotler et al., 2009) provide a conserva- tive precis of the Lepidoziaceae and do not propose a structure below family level. Crandall-Stotler et al. (2009) noted in their review of liverwort classification, based on a decade of phyloge- netic analyses, that broader sampling within the Lepidoziaceae would be necessary before a revision of its internal classification could be presented. The most recent and complete subfamilial classification is that of Schuster (2000), an account that largely fol- lows Schuster (1969), but places Drucella integristipula, Neogrollea notabilis and Protocephalozia ephemeroides in monotypic subfami- lies. In the absence of robust modern phylogenetic hypotheses, extensive taxonomic study of the family has not provided stable subfamily, generic or subgeneric delimitations. For example, the monotypic genus Neogrollea was recently removed from the Lepidoziaceae to its own monotypic suborder Neogrollineae (Engel and Braggins, 2001) and Megalembidium insulanum was transferred from Lepidozioideae to a monotypic Megalembidioideae (Engel and Braggins, 2005). The delimitation of genera has followed often geographically restricted traditional morphological treatments and as a result generic limits remain fluid (Engel and Merrill, 1994, 2004; Engel and Schuster, 2001; Evans, 1934; Grolle, 1964; Hodgson, 1955; Schuster, 1980). 1055-7903/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved. doi:10.1016/j.ympev.2011.02.006 ⇑ Corresponding author. Address: Rm 409 Heydon-Laurence Building (A08), School of Biological Sciences, University of Sydney, NSW 2006, Australia. Fax: +61 2 9351 4119. E-mail address: endymion.cooper@sydney.edu.au (E.D. Cooper). Molecular Phylogenetics and Evolution 59 (2011) 489–509 Contents lists available at ScienceDirect Molecular Phylogenetics and Evolution journal homepage: www.elsevier.com/locate/ympev