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University of Florida Thesis Or Dissertation Formatting Template GENETIC VARIATION IN CALADIUM AND PETUNIA By ZHE CAO 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 2016 © 2016 ZHE CAO To my family and friends ACKNOWLEDGMENTS I would like to thank my Ph.D. committees, Drs. Zhanao Deng, David G. Clark, Kenneth Quesenberry, Natalia Peres, and Ryan Warner for their guidance on my research projects. We are grateful to Dr. Jaroslav Doležel (Institute Experimental Botany, Olomouc, Czech Republic) for providing rye seeds and to the University of Florida’s Interdisciplinary Center for Biotechnology Research for access to its flow cytometer. Completion of my Ph.D. projects required five years. I also must thank my parents for supporting me and understanding my absence from their lives for such a long time. My wife always provides me with supports and advices both in daily life and my research projects and without which I would not finish my projects. 4 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 9 LIST OF FIGURES ........................................................................................................ 11 LIST OF ABBREVIATIONS ........................................................................................... 13 ABSTRACT ................................................................................................................... 15 CHAPTER 1 LITERATURE REVIEW .......................................................................................... 17 Overview ................................................................................................................. 17 Floriculture in Florida ........................................................................................ 17 Caladium .......................................................................................................... 18 Petunia ............................................................................................................. 20 Challenges in Caladium and Petunia Breeding ................................................ 21 Exploration of Genetic Variation ............................................................................. 23 Use of Wild Relatives, Progenitor Species, and Primitive Cultivars .................. 23 Inducing Genetic Variation (Somaclonal Variation) .......................................... 24 Genetic Variation in Caladium ................................................................................ 26 Caladium Wild Species..................................................................................... 26 Somaclonal Variation ....................................................................................... 27 Variation in Major Leaf Characteristics ............................................................. 28 Genetic Variation in Petunia ................................................................................... 29 Important Wild Species in Petunia .................................................................... 29 Challenges in Using Wild Germplasm in Breeding Programs ........................... 30 Linkage Map Development and QTL Analysis in Petunia Wild Germplasm ..... 30 Research Objectives ............................................................................................... 31 Caladium .......................................................................................................... 31 Petunia ............................................................................................................. 32 2 INTERSPECIFIC GENOME SIZE AND CHROMOSOME NUMBER VARIATION IN CALADIUM (ARACEAE) .................................................................................... 33 Introduction ............................................................................................................. 33 Materials and Methods............................................................................................ 34 Plant Materials .................................................................................................. 34 Genome Size Determination ............................................................................ 35 Chromosome Counting .................................................................................... 36 Results .................................................................................................................... 37 Genome Size Estimates ................................................................................... 37 5 Chromosome Counts ....................................................................................... 39 Discussion .............................................................................................................. 41 Classification of Caladium Species .................................................................. 41 Genome Size Variation in Caladium ................................................................. 43 Chromosome Number Variation in Caladium ................................................... 44 Possible Modes of Genome Size and Chromosome Number Evolution in Caladium ....................................................................................................... 47 Implications for Preservation of Caladium Diversity and Caladium Breeding ... 50 3 SOMACLONAL VARIATION IN CALADIUM ’RED FLASH’ .................................... 62 Introduction ............................................................................................................. 62 Materials and Methods............................................................................................ 63 Plant Materials .................................................................................................. 63 Tissue Culture and Plant Regeneration ............................................................ 64 Plants Acclimatization ...................................................................................... 64 Identification of Somaclonal Variants ............................................................... 65 Determination of Nuclear DNA Contents .......................................................... 66 Data Analysis ................................................................................................... 66 Chromosome Counting .................................................................................... 67 SSR Marker Analysis ....................................................................................... 67 Results .................................................................................................................... 68 Effect of PGR and Explant Type on Plant Regeneration and Occurrence of Somaclonal Variation .................................................................................... 68 Somaclonal Variation and Variant Types ......................................................... 69 Nuclear DNA Contents ..................................................................................... 71 Chromosome Counts ....................................................................................... 72 SSR Marker Profiling ........................................................................................ 73 Discussion .............................................................................................................. 74 4 INHERITANCE OF LEAF RUGOSITY IN CALADIUM ............................................ 86 Introduction ............................................................................................................. 86 Materials and Methods............................................................................................ 87 Plant Materials .................................................................................................. 87 Flower Induction and Crossing ......................................................................... 88 Progeny Growing .............................................................................................. 88 Phenotypic Analysis ......................................................................................... 89 Statistical Analysis ............................................................................................ 89 Results .................................................................................................................... 90 Inheritance of Rugose Leaf .............................................................................. 90 Genetic Relationship between Rugose Leaf and Leaf Shape .......................... 91 Genetic Relationship between Rugose Leaf and Leaf Spotting ........................ 92 Genetic Relationship between Rugose Leaf and Main Vein Color ................... 94 Genetic Distances and Gene Order among Rugose Leaf, Leaf Spotting and Main Vein Color ............................................................................................. 95 Discussion .............................................................................................................. 96 6 5 INHERITANCE OF LEAF BACKGROUND COLOR IN CALADIUM ..................... 111 Introduction ..........................................................................................................
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