GENETIC STERILIZATION AND REPRODUCTIVE BIOLOGY OF LANTANA CAMARA By DAVID MARK CZARNECKI II 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 2011 1 © 2011 David Mark Czarnecki II 2 To my family and friends 3 ACKNOWLEDGMENTS Completion of this project required the help of numerous individuals over the last few years. I would like to thank my parents for understanding my absence from their lives and my inability to participate in the important events that I have missed because of the commitments associated with my degree. My friends over the years have provided me with support, advice, and much needed grounding without which I would not have had the focus to complete my projects. Lastly I would like to thank my professor and advisors, who have given me the guidance and advice to learn and grow as a scientist. 4 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES .......................................................................................................... 10 LIST OF FIGURES ........................................................................................................ 12 LIST OF ABBREVIATIONS ........................................................................................... 13 ABSTRACT ................................................................................................................... 15 CHAPTER 1 INTRODUCTION .................................................................................................... 17 Overview and Rationale .......................................................................................... 17 Floriculture Industry and Concerns about Introduced Plant Species ................ 17 Lantana camara ............................................................................................... 18 Need To Sterilize Lantana camara ................................................................... 19 Invasive Plants ........................................................................................................ 22 Potential Ecological and Economic Damages .................................................. 22 Managing Invasive Plants ................................................................................. 23 Regulatory Measures ....................................................................................... 24 Biological Factors Determining Invasive Potential of Plants ................................... 26 Seed Production and Dispersal ........................................................................ 26 Pollen Production and Dispersal ...................................................................... 26 Vegetative Propagules ..................................................................................... 27 Assessing Invasive Potential of Plants ............................................................. 27 Genetic Approaches to Reduce or Eliminate Invasive Potential of Plants .............. 28 Traditional Breeding ......................................................................................... 29 Mutational Breeding ......................................................................................... 30 Transgenic Approaches.................................................................................... 30 Ploidy Manipulation .......................................................................................... 31 Research Objectives ............................................................................................... 31 2 MALE FERTILITY OF LANTANA CAMARA ........................................................... 33 Rationale ................................................................................................................. 33 Materials and Methods............................................................................................ 34 Plant Materials .................................................................................................. 34 Ploidy Analysis ................................................................................................. 35 Pollen Staining ................................................................................................. 36 Experimental Designs ...................................................................................... 37 Statistical Analysis ............................................................................................ 37 Meiotic Observation .......................................................................................... 38 5 Results and Discussion........................................................................................... 38 Ploidy Analysis ................................................................................................. 38 Dye Comparison for Lantana Pollen Stainability Determination ....................... 39 Pollen Stainability of L. camara Cultivars and Breeding Lines .......................... 39 Diploids ...................................................................................................... 40 Triploids ..................................................................................................... 41 Tetraploids ................................................................................................. 41 Pentaploids and hexaploids ....................................................................... 42 Meiotic Abnormalities ....................................................................................... 42 Summary ................................................................................................................ 43 3 FEMALE FERTILITY OF LANTANA CAMARA ....................................................... 51 Rationale ................................................................................................................. 51 Materials and Methods............................................................................................ 53 Plant Materials .................................................................................................. 53 Assessing Fruit Production ............................................................................... 53 Insect Damage ................................................................................................. 54 Evaluating Seed Germination ........................................................................... 55 Calculating Female Fertility Index .................................................................... 55 Plant Dry Weight .............................................................................................. 55 Experimental Design ........................................................................................ 56 Statistical Analysis ............................................................................................ 56 Results and Discussion........................................................................................... 56 Fruit Production in L. camara ........................................................................... 56 Seed Germination in L. camara ........................................................................ 57 Female Fertility Index (FFI) of L. camara.......................................................... 58 Fruit Production, Seed Germination and Female Fertility Index of Diploid L. camara .......................................................................................................... 58 Fruit Production, Seed Germination and Female Fertility Index of Triploid L. camara .......................................................................................................... 59 Non-UFG-producing triploids ..................................................................... 59 UFG-producing triploids ............................................................................. 60 Fruit Production, Seed Germination and Female Fertility Index of Tetraploid L. camara ...................................................................................................... 61 Non-UFG-producing tetraploids ................................................................. 61 UFG-producing tetraploids ......................................................................... 61 Fruit Production, Seed Germination and Female Fertility Index of Pentaploid and Hexaploid L. camara .............................................................................. 62 Correlation Analysis ......................................................................................... 63 Principal Component Analysis .......................................................................... 64 Summary ................................................................................................................ 64 4 OCCURRENCE OF UNREDUCED FEMALE GAMETES LEADS TO SEXUAL POLYPLOIDIZATION IN LANTANA CAMARA ....................................................... 72 Rationale ................................................................................................................. 72 6 Materials and Methods............................................................................................ 75 Plant Materials .................................................................................................. 75 Pollinations