DISSERTATION Titel der Dissertation Molecular phylogeny, evolution and biogeography of Ranunculus (Ranunculaceae) and related genera Verfasserin Khatere Emadzade (MSc) angestrebter akademischer Grad Doktorin der Naturwissenschaften (Dr. rer. nat.) Matrikelnummer: 0549709 Studienkennzahl lt. Studienblatt: A091 438 Dissertationsgebiet lt. Studienblatt: Botanik Betreuer: Doz. Dr. Elvira Hörandl Wien, im Jänner 2010 Table of contents Acknowledgment …(iii) Co-authorship statement …(iv) Abstract …(v) Zusammenfassung …(vii) Chapter 1: General Introduction …(1) Chapter 2: A molecular phylogeny, morphology and classification of genera of Ranunculeae (Ranunculaceae) …(11) Chapter 3: Northern Hemisphere origin, transoceanic dispersal, and diversification of Ranunculeae (Ranunculaceae) in the Tertiary …(52) Chapter 4: The biogeographical history of the cosmopolitan genus Ranunculus L. (Ranunculaceae) in the temperate to meridional zones …(83) Chapter 5: Rapid speciation in high alpine and arctic species of Ranunculus during the Quaternary …(139) Appendixes (abstracts of contributions to international conferences) …(171) Curriculum Vitae …(178) ii Acknowledgement I am thankful to my supervisor, Doz. Dr. Elvira Hörandl, whose encouragement, guidance and support from the initial to the final level enabled me to develop an understanding of the subject. I gratefully thank Prof. Dr. Tod Stuessy and Prof. Dr. Rose Samuel for their outstanding teaching and instruction. Deepest gratitude is also due to Dr. Karin Tremetsberger, Dr. Hanna Weiss-Schneeweiss, Dr. Gerald Schneeweiss, and Dr. Herman Voglmayr, who answered my questions with patience. Many thanks go in particular to Dr. Carolin Anna Rebernig for her wonderful friendship during my stay in Vienna and assisting me in various ways. I must thank my colleagues Mag. Michael Barfuss, Mag. Cordula Blöch and Mag. Anne-Caroline Cosendai for their support ranging from practical suggestions to the analyses. Ing. Elfriede Grasserbauer, Verena Klejna, and Mag. Gudrun Kohl are thanked for their laboratory advice and their willingness to share their experience with me. Mag. Jan Rodewald is thanked for his great support in the lab. Many thanks go in particular to Anton Russell, Patricio Lopez, Mag. Katharina Bardy, Mag. Stephan Safer, and Dr. Pedro Escobar for giving me such a pleasant time when working together with them. I would like to thank the members of the department of structural and functional Botany in particularly DDr. Heidemarie Halbritter. I thank all members of the Institute of Botany of the University of Vienna for their kind reception through these years. I grateful to the curators of the herbaria BISH, CAN, CONN, GB, LD, LE, LI, M, MPN, RM, TARI, VALD, WU, W, ZH, ZT for the loan of herbarium specimens and permission to use materials for DNA extractions. My sincere thanks also go to my colleagues at the Herbarium of Ferdowsi University of Mashhad (FUMH) for their support. I offer my regards and blessings to all of those who collected material from different continents and this thesis could not have been accomplished without their assistance. The author would also like to acknowledge a PhD student grant of the Austrian Exchange Service (ÖAD), the Commission for Interdisciplinary Ecological Studies (KIÖS) of the Austrian Academy of Sciences (ÖAW) for providing the financial means. I would like to thank my family, especially my parents for their warm-hearted support and understanding my love to research. Words fail me to express my appreciation to my husband Dr. Alireza Emami-Nouri. Alireza inspired me to my best times and carried me through my most difficult times. iii Co-authorship statement Chapters 2, 3, 4, and 5 were prepared as papers in international journals. For all of the studies I developed the idea and the methodology. I collected ca. 60 new samples in Iran and the Himalayas, prepared all of the molecular data, performed the analyses and wrote the manuscripts. Dr. Elvira Hörandl contributed significantly to project design, interpretation of the results and editing of the manuscripts. Chapter 2 (Paper 1): was accepted as a manuscript in Taxon co-authored with Dr. Elvira Hörandl, Prof. Peter Lockhart (Massey University, New Zealand) and Dr. Carlos Lehnebach (Museum of New Zealand, New Zealand). Dr. Lockhart contributed to the experimental design and helpful discussion particularly in Neighbor Net analysis. Dr. Lehnebach provided some materials, valuable suggestions and he did the histological part. Chapter 3 (Paper 2): was submitted as a manuscript to Journal of Biogeography authored by only Dr. Elvira Hörandl and me. Chapter 4 (Paper 3): was submitted as a manuscript to Molecular Phylogenetics and Evolution co-authored with Dr. Elvira Hörandl, Prof. H. Peter Linder (University of Zurich, Switzerland) and Dr. Berit Gehrke (University of Cape Town, South Africa). Prof. Linder provided directional advice and Dr. Berit Gehrke prepared sequences of African samples, edited the manuscript and provided directional advice. Chapter 5 (Paper 4): was prepared as a manuscript to Evolution co-authored with Dr. Elvira Hörandl, Dr. Matthias Hoffmann (Martin-Luther-Universität Halle-Wittenberg, Germany), and Dr. Natalia Tkach (Martin-Luther-Universität Halle-Wittenberg, Germany). Dr. Matthias Hoffmann provided advice on statistical analysis, and contributed to manuscript preparation. Dr. Natalia Tkach prepared sequences of Arctic species. iv Abstract The thesis aims at a reconstruction of the phylogeny and biogeographical history of Ranunculeae, with a focus on Ranunculus s.str. Ranunculus s.str. is a cosmopolitan genus with approximately 600 species and the largest genus in Ranunculaceae. Ranunculus is distributed on all continents and it has a worldwide distribution from the Tropics to the arctic and subantarctic zones. A molecular phylogenetic analysis based on nuclear and plastid markers (ITS, matK/trnK, psbJ-petA) provides the framework for understanding relationships, biogeography and character evolution in Ranunculus and related genera. Combined molecular data of c. 240 species from all continents reveal a large core clade comprising Ranunculus s.str., excluding the small genera Laccopetalum, Krapfia, Ceratocephala, Myosurus, Ficaria, Coptidium, Beckwithia, Cyrtorhyncha, Halerpestes, Peltocalathos, Callianthemoides, and Arcteranthis, but including the water-buttercups and the monotypic genus Aphanostemma. Molecular and morphological data reveal that a classification accepting several small genera and one big genus Ranunculus s.str. reflects best the phylogeny and the morphological diversity of the tribe. Age estimates based on molecular dating suggest that Ranunculeae diversified between the late Eocene and the late Miocene. Biogeographical analysis suggests a northern hemispheric origin of the tribe and multiple colonization of the S. hemisphere. Results of biogeographical analyses of Ranunculus s.str. support multiple colonizations of all continents. Dispersals between continents must have occurred via migration over land bridges, or via long distance dispersal. In southern Eurasia, isolation of the western Mediterranean and the Caucasus region during the Messinian was followed by range expansions and speciation in both areas. In the Pliocene and Pleistocene, radiations happened independently in the summer-dry W. Mediterranean-Makaronesian and in the E. Mediterranean-Irano-Turanian regions, with three independent shifts to alpine humid climates in the Alps and in the Himalayas. In previously glaciated areas, rapid colonization was followed by speciation, and regional radiations. This pattern is seen in a clade comprising arctic, Central Asian, North American and European lowland taxa. In North America, the availability of a large area and a broad range of habitats triggered allopatric speciation and adaptive radiation. In contrast, in the Himalayas, the alpine species are restricted to a narrow ecological zone in high altitudes, v resulting in extensive hybridization. The Arctic was colonized multiple times without a pronounced radiation. Altogether, the success of Ranunculus can be referred to a high ability not only to long- distance dispersal to new areas but also to rapid speciation. vi Zusammenfassung Diese Dissertation hat zum Ziel, eine Rekonstruktion der Phylogenie und der Biogeographie der Ranunculeae mit dem Schwerpunkt auf der Gattung Ranunculus s. str. zu erstellen. Ranunculus s.str ist eine kosmopolitische, von tropischen bis in arktische Gebiete verbreitete Gattung und mit ca. 600 Arten die größte innerhalb der Ranunculaceae. Die in dieser Arbeit durchgeführte molekular-phylogenetische Untersuchung von Ranunculus, basierend Sequenzen der Kern- und Chloroplasten-DNS (ITS, matK/trnK, psbJ-petA), bietet die Basis für das Verständnis von Verwandtschaftsverhältnissen, Biogeographie und Evolution innerhalb der Gattung genauso wie zu verwandten Gattungen. Die Kombination dieser molekularen Daten von ca. 240 Arten, gesammelt über das gesamte Verbreitungsgebiet der Tribus. resultierte in einer großen Klade, die Ranunculus s.str. beinhaltet, jedoch Laccopetalum, Krapfia, Ceratocephala, Myosurus, Ficaria, Coptidium, Beckwithia, Cyrtorhyncha, Halerpestes, Peltocalathos, Callianthemoides und Arcteranthis exkludiert. Morphologische Untersuchungen unterstützen die Schlussfolgerung, dass eine Gliederung in mehrere kleine Gattungen und eine große Gattung Ranunculus s.str. die phylogenetischen Verhältnisse sowie die morphologische Diversität innerhalb der Tribus bestmöglich reflektiert.