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SYSTEMATICS OF TRIBE TRICHOCEREEAE AND POPULATION GENETICS OF Haageocereus (CACTACEAE) By MÓNICA ARAKAKI MAKISHI A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2008 1 © 2008 Mónica Arakaki Makishi 2 To my parents, Bunzo and Cristina, and to my sisters and brother. 3 ACKNOWLEDGMENTS I want to express my deepest appreciation to my advisors, Douglas Soltis and Pamela Soltis, for their consistent support, encouragement and generosity of time. I would also like to thank Norris Williams and Michael Miyamoto, members of my committee, for their guidance, good disposition and positive feedback. Special thanks go to Carlos Ostolaza and Fátima Cáceres, for sharing their knowledge on Peruvian Cactaceae, and for providing essential plant material, confirmation of identifications, and their detailed observations of cacti in the field. I am indebted to the many individuals that have directly or indirectly supported me during the fieldwork: Carlos Ostolaza, Fátima Cáceres, Asunción Cano, Blanca León, José Roque, María La Torre, Richard Aguilar, Nestor Cieza, Olivier Klopfenstein, Martha Vargas, Natalia Calderón, Freddy Peláez, Yammil Ramírez, Eric Rodríguez, Percy Sandoval, and Kenneth Young (Peru); Stephan Beck, Noemí Quispe, Lorena Rey, Rosa Meneses, Alejandro Apaza, Esther Valenzuela, Mónica Zeballos, Freddy Centeno, Alfredo Fuentes, and Ramiro Lopez (Bolivia); María E. Ramírez, Mélica Muñoz, and Raquel Pinto (Chile). I thank the curators and staff of the herbaria B, F, FLAS, LPB, MO, USM, U, TEX, UNSA and ZSS, who kindly loaned specimens or made information available through electronic means. Thanks to Carlos Ostolaza for providing seeds of Haageocereus tenuis, to Graham Charles for seeds of Blossfeldia sucrensis and Acanthocalycium spiniflorum, to Donald Henne for specimens of Haageocereus lanugispinus; and to Bernard Hauser and Kent Vliet for aid with microscopy. I would like to express my gratitude to current and former members of the Soltis Lab, FLAS Herbarium, and Florida Museum of Natural History, particularly Matt Gitzendanner, Heather Loring, Pablo Speranza, Jennifer Tate, Vaughan Symonds, Luiz de Oliveira, Mi-Jeong Yoo, Sam Brockington, Evgeny Mavrodiev, Summer Scobell, Christine Edwards, Josh Clayton, 4 Lucas Majure, Claudia Segovia, Lorena Endara, Mark Whitten, Kent Perkins, Andres Lopez and Tuuli Makinen. My deepest thanks to my old and new friends, for their patience and encouragement at all times. This work was supported by grants from the American Society of Plant Taxonomy (ASPT), the Botanical Society of America (BSA), the Cactus and Succulent Society of America (CSSA), the International Association for Plant Taxonomy (IAPT), and the National Science Foundation (NSF DEB-0608273). 5 TABLE OF CONTENTS page ACKNOWLEDGMENTS ...............................................................................................................4 LIST OF TABLES...........................................................................................................................8 LIST OF FIGURES .........................................................................................................................9 ABSTRACT...................................................................................................................................11 CHAPTER 1 INTRODUCTION: SYSTEMATIC STATUS OF THE CACTACEAE, TRIBE TRICHOCEREEAE, AND GENUS Haageocereus ..............................................................13 Cactaceae ................................................................................................................................13 Tribe Trichocereeae (Subfamily Cactoideae).........................................................................16 Haageocereus Backeb. ...........................................................................................................18 2 SYSTEMATICS OF TRIBE TRICHOCEREEAE AND GENUS Haageocereus ................24 Introduction.............................................................................................................................24 Materials and Methods ...........................................................................................................30 Taxon Sampling...............................................................................................................30 DNA Isolation, Amplification, and Sequencing..............................................................31 Alignment ans Phylogenetic Analysis.............................................................................34 Results.....................................................................................................................................36 Descriptive Data ..............................................................................................................36 Parsimony Analysis.........................................................................................................36 Maximum Likelihood Analysis.......................................................................................38 Bayesian Analysis ...........................................................................................................39 Discussion...............................................................................................................................40 Phylogenetic Inference ....................................................................................................40 Phylogenetic Relationships in Trichocereeae..................................................................41 Phylogenetic Relationships in Haageocereus .................................................................48 Radiation of Trichocereeae..............................................................................................49 Conclusions.............................................................................................................................50 3 DEVELOPMENT OF POLYMORPHIC MICROSATELLITE LOCI FOR THE EXAMINATION OF GENETIC DIVERSITY IN Haageocereus ........................................65 Introduction.............................................................................................................................65 Materials and Methods ...........................................................................................................66 Results and Discussion ...........................................................................................................67 6 4 CHROMOSOME COUNTS AND EVIDENCE OF POLYPLOIDY IN Haageocereus AND OTHER TAXA OF TRIBE TRICHOCEREEAE.........................................................72 Introduction.............................................................................................................................72 Materials and Methods ...........................................................................................................73 Results and Discussion ...........................................................................................................74 5 CLONAL REPRODUCTION AND APOMIXIS IN Haageocereus.....................................82 Materials and Methods ...........................................................................................................84 Examination of Apomixis................................................................................................85 Microsatellite Amplification and Analysis......................................................................85 Results and Discussion ...........................................................................................................86 Agamospermy..................................................................................................................86 Genetic Diversity.............................................................................................................86 Implications for Conservation .........................................................................................88 6 EXAMINATION OF RETICULATE EVOLUTION BETWEEN Haageocereus AND Espostoa..................................................................................................................................91 Introduction.............................................................................................................................91 Materials and Methods ...........................................................................................................96 Plant Sampling.................................................................................................................96 Nuclear Microsatellite Data.............................................................................................96 Chloroplast DNA Data ....................................................................................................97 Results and Discussion ...........................................................................................................98 7 CONCLUDING REMARKS................................................................................................108 REFERENCES ............................................................................................................................111 BIOGRAPHICAL SKETCH .......................................................................................................125 7 LIST OF TABLES Table page 1-1 Classification