MOLECULAR SYSTEMATICS AND ANATOMY OF VANDEAE (ORCHIDACEAE): THE EVOLUTION OF MONOPODIAL LEAFLESSNESS By BARBARA S. CARLSWARD 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 2004 Copyright 2004 by Barbara S. Carlsward ACKNOWLEDGMENTS I would like to thank my advisor, William Louis Stern, for his invaluable guidance. He welcomed me into his lab at the beginning of my botany career and has continually nurtured my love of plants and their anatomy. I would also like to thank W. Mark Whitten, who has been a friend and mentor. Without his constant guidance, I would have failed miserably in the molecular laboratory. During the course of my doctoral degree, Norris H. Williams has offered useful counsel, allowed me unlimited access to his laboratory, and given me office space to write my dissertation. From the beginning of my botany career, Walter S. Judd has been instrumental in developing my love for plants and has always given helpful advice in the process of delimiting anatomical data for cladistic analysis. As a committee member, Robert J. Ferl provided insightful advice and guidance. Robert L. Dressler was always willing to help me identify orchids and gave helpful advice on tricky taxonomic issues. James D. Ackerman was also extremely generous with his taxonomic advice, especially regarding Dendrophylax. Gustavo Romero (AMES Herbarium, Harvard University, Massachusetts) provided assistance in finding the taxonomic articles necessary for the nomenclatural transfers of Solenangis. Alec M. Pridgeon has continually been willing to offer guidance and counsel, most notably with the interpretation of the vandaceous anatomy. Lars Jonsson was kind enough to donate his iii monographic work on Microcoelia and give advice on leafless Vandeae. Gunnar Seidenfaden, now deceased, graciously sent me his last copy of Opera Botanica 95 (“Orchid genera in Thailand XIV. Fifty-nine vandoid genera”). Eric A. Christenson provided sage taxonomic advice and was always willing to identify specimens of Aeridinae. I also thank Sherwin Carlquist and Edward L. Schneider for providing the seed of an idea for this project. I am gratefule to Jennifer A. Thorsch and Bruce H. Tiffney for their friendship and mentorship during my brief graduate career at the University of California, Santa Barbara. I would like to acknowledge the tremendous generosity of the following commercial and hobbyist orchid growers: Isobyl La Croix (Uzumara Orchids, Gairloch, Scottland); Robert Fuchs (R. F. Orchids, Inc., Homestead, Florida); Martin Motes (Motes Orchids, Homestead, Florida); Claude Hamilton (Hamlyn Orchids, Kingston, Jamaica); Gaspar Silvera (Orquideas Tropicales, Panama City, Panama); William A. (Andy) Phillips (Andy’s Orchids, Encinitas, California); Jim Rose (Cal-Orchid, Santa Barbara, California); Erich Michel (Hoosier Orchid Company, Indianapolis, Indiana); Rick Reese (Rick’s Tropica, New Port Richey, Florida); Countryside Orchids (Corrales, New Mexico); Johan Hermans (Royal Botanic Gardens, Kew, London, England); James D. Ackerman (University of Puerto Rico, Puerto Rico); Germán Carnevali (Centro de Investigación Científica de Yucatán, Mexico); Paul Simon (private grower, Los Altos Hills, California); and James Watts (private grower, Hollywood, Florida). Harry Luther, Bruce Holst, and Wesley Higgins of The Marie Selby Botanical Gardens were all extremely iv generous with their resources and time. I also acknowledge the help of Paula J. Rudall, Mark W. Chase, Edith Kapinos, and the late Timothy Lawrence at The Royal Botanic Gardens, Kew, for their help in processing, vouchering, and shipping the anatomical and DNA specimens donated by Johan Hermans. Benny Bytebier (University of Stellenbosch, South Africa) collected and vouchered most of my angraecoid specimens from Africa, for which I am thankful. Assistance and use of equipment provided by the Electron Microscopy Core Laboratory at the University of Florida (especially Karen L. Kelley) were essential to the success of my anatomical work. I would like to thank three of my fellow graduate students for their assistance in completing this project: Mario A. Blanco collected Campylocentrum brenesii from Costa Rica, Kurt M. Neubig sequenced several of my last DNA specimens, and J. Michael Heaney lent me some of his Polystachyinae slides for anatomical observation. I want to also thank the helpful staff of the Department of Botany and the University of Florida Herbarium. This research was supported by the National Science Foundation (Doctoral Dissertation Improvement Grant, DEB-0104566), the Lewis and Varina Vaughn Fellowship in Orchid Biology, the American Orchid Society’s 11th World Orchid Conference Fellowship, and the College of Liberal Arts and Sciences Dissertation Fellowship. William Stern provided funds to complete much of the molecular and anatomical work, and Norris Williams freely allowed me to use the supplies and equipment in his molecular laboratory. v I am grateful to my grandparents, Carl and Ellen Carlsward, who have supported and loved me unconditionally throughout my life. I would also like to thank my life-long partner and friend, J. Richard Abbott. His love, guidance, and infectious passion for plants have helped me grow throughout my botanical career. vi TABLE OF CONTENTS Page ACKNOWLEDGMENTS .......................................................................................iii LIST OF TABLES................................................................................................. ix LIST OF FIGURES ............................................................................................... x ABSTRACT.........................................................................................................xvi CHAPTER 1 INTRODUCTION............................................................................................1 Systematics ....................................................................................................2 Leaflessness ................................................................................................11 Anatomy .......................................................................................................13 Objectives.....................................................................................................14 Methods........................................................................................................14 2 MOLECULAR PHYLOGENY OF VANDEAE................................................16 Introduction...................................................................................................16 Materials and Methods .................................................................................17 Plant Material .........................................................................................17 Extraction ...............................................................................................28 Amplification...........................................................................................30 Sequencing ............................................................................................32 Data Analysis .........................................................................................33 Results .........................................................................................................36 ITS Analyses of Vandeae.......................................................................36 ITS and Chloroplast Analyses of Angraecinae and Aerangidinae..........45 ITS matrix ........................................................................................45 matK matrix .....................................................................................46 trnL-F matrix ....................................................................................51 Combined chloroplast matrix ...........................................................56 Combined ITS and chloroplast matrix..............................................60 Discussion ....................................................................................................65 vii 3 VEGETATIVE ANATOMY OF VANDEAE ....................................................78 Introduction...................................................................................................78 Material and Methods...................................................................................87 Results .......................................................................................................113 Anatomy of Tribe Vandeae ..................................................................113 Anatomy of Subtribe Angraecinae .................................................118 Anatomy of Subtribe Aeridinae ......................................................196 Anatomy of Tribe Epidendreae, Subtribe Polystachyinae ....................207 Phylogenetic Analyses of Vandeae......................................................212 Discussion ..................................................................................................213 Comparative Anatomy..........................................................................213 Cladistics..............................................................................................221 4 ANATOMICAL EVOLUTION WITHIN ANGRAECINAE..............................223 Introduction.................................................................................................223