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University of Florida Thesis Or Dissertation IN VITRO ECOLOGY OF Calopogon tuberosus var. tuberosus: A NEW CONCEPT IN SPECIES CONSERVATION By PHILIP KAUTH 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 2009 1 © 2009 Philip Kauth 2 To Meredith, who else… 3 ACKNOWLEDGMENTS I thank Dr. Michael Kane for his dedication, passion, enthusiasm, and encouragement, which in turn fueled my passion and excitement. I thank my co-chair, Dr. Wagner Vendrame for his support and encouragement from a distance. I also thank Dr. Carrie Reinhardt-Adams, Dr. Debbie Miller, and Dr. Thomas Sheehan for serving on my supervisory committee and sharing their knowledge, ideas, and excitement. Over the last several years I had the opportunity to share a work environment with great people. I could not have shared space, ideas, and knowledge over the years with better colleagues and friends than Tim Johnson, Daniela Dutra, Scott Stewart, Xiuli Shen, and Carmen Valero- Aracama. I especially thank Tim for the hours of brainstorming, talking, watching the fights, and trips to the Refuge. I thank Nancy Philman not only for her incredible friendship, but also for much of my development (even though she probably will deny her involvement). This research could not have been conducted without the assistance from many that have either collected seed, issued permits, served as field trip guides, or provided thoughtful insights including Larry Richardson (USFWS-FPNWR), Mary Bunch (South Carolina Heritage Preserves-SCDNR), Jim Fowler (Greenville, South Carolina), Howard Lorenz (Wisconsin), and Doug Goldman (Harvard University). I thank Kip Knudson not only for collecting seed and leading me through Carney Fen numerous times, but also for influencing me more than me takes credit for. I also thank Dr. Hector Perez and Dr. Charles Guy for use of their incubators and growth chambers. I thank my parents for their endless support, love, and encouragement throughout my LONG educational process. I thank my two beautiful dogs, Mya and Ladybug, for providing hours of laughter, and for showing me that life is a true blessing and that everyone deserves a second chance at life. Finally I thank the love of my life, Meredith. I could not have done this without her love, sacrifice, and support and for that I am forever grateful. 4 TABLE OF CONTENTS page ACKNOWLEDGMENTS ...............................................................................................................4 LIST OF TABLES...........................................................................................................................9 LIST OF FIGURES .......................................................................................................................10 ABSTRACT...................................................................................................................................13 CHAPTER 1 LITERATURE REVIEW .......................................................................................................15 Introduction.............................................................................................................................15 The Orchid Seed .....................................................................................................................16 Orchid Seed Germination .......................................................................................................18 History and Background..................................................................................................18 Symbiotic Orchid Seed Germination...............................................................................19 Asymbiotic Orchid Seed Germination ............................................................................20 Factors Effecting Orchid Seed Germination ...................................................................25 Photoperiod ..............................................................................................................25 Temperature .............................................................................................................28 Ecotypes..................................................................................................................................31 Introduction .....................................................................................................................31 Importance of Ecotypes to Plant Conservation ...............................................................32 Ecotype Development and Differentiation......................................................................34 Common garden studies...........................................................................................34 Seed germination ecology ........................................................................................37 Biomass allocation ...................................................................................................38 In vitro ecology ........................................................................................................40 Ecotypic Differentiation in Orchids ................................................................................42 Seed source...............................................................................................................42 Pollination ecotypes .................................................................................................44 Plant of Study .........................................................................................................................45 Diversity within Calopogon tuberosus............................................................................46 Floral Biology of Calopogon tuberosus..........................................................................48 Seed Germination of Calopogon tuberosus ....................................................................49 Habitat Descriptions of Calopogon tuberosus........................................................................51 Carney Fen.......................................................................................................................52 Ashmore Heritage Preserve.............................................................................................54 Site C ...............................................................................................................................55 Eva Chandler Heritage Preserve......................................................................................56 Goethe State Forest..........................................................................................................58 5 Florida Panther National Wildlife Refuge.......................................................................60 2 EFFECTS OF PHOTOPERIOD AND GERMINATION MEDIA ON IN VITRO SEED ECOLOGY OF Calopogon tuberosus....................................................................................70 Introduction.............................................................................................................................70 Materials and Methods ...........................................................................................................71 Seed Source .....................................................................................................................71 Seed Viability Test ..........................................................................................................72 Media and Seed Preparation............................................................................................72 Photoperiod Effects on Asymbiotic Germination and Early Seedling Development .....73 Photoperiod Effects on Advanced In Vitro Seedling Development................................73 Asymbiotic Germination Media Evaluation....................................................................74 Soil Analysis....................................................................................................................75 Statistical Analysis ..........................................................................................................75 Results.....................................................................................................................................76 Seed Viability ..................................................................................................................76 Photoperiod Effects on Germination and Early Development ........................................76 Photoperiod effects on Advanced Seedling Development ..............................................78 Media Effects on Germination and Early Development .................................................79 Media Effects on Advanced Seedling Development.......................................................81 Soil Nutrient Analysis .....................................................................................................81 Discussion...............................................................................................................................82 Seed Viability and Quality ..............................................................................................82 Photoperiod......................................................................................................................83 Media Screen and Soil Nutrient Availability ..................................................................85
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