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EVOLUTIONARY PATTERNS in the DILATATA GROUP (Paspalum, POACEAE): a POLYPLOID/AGAMIC COMPLEX

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EVOLUTIONARY PATTERNS in the DILATATA GROUP (Paspalum, POACEAE): a POLYPLOID/AGAMIC COMPLEX EVOLUTIONARY PATTERNS IN THE DILATATA GROUP (Paspalum, POACEAE): A POLYPLOID/AGAMIC COMPLEX By PABLO R. SPERANZA 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 2005 Copyright 2005 by Pablo R. Speranza This dissertation is dedicated to my son Mauricio who never forgot to give me a hug when I left home for work at night. ACKNOWLEDGMENTS I want to thank my advisor, Pamela Soltis, for having always been available and willing to help and provide support during my whole program. I also wish to thank my committee members (Pamela Soltis, Douglas Soltis, Kenneth Quesenberry, and Gloria Moore) for having made of each meeting a constructive and worthy experience, each providing a different point of view on my work. I am indebted to Enrique Estramil and Marcos Malosetti for their help with data analysis, and to my laboratory mates, particularly Joshua Clayton, Vaughan Symonds, and Jennifer Tate for their invaluable help in writing this dissertation. I also want to thank Gabriel Rua, Rodrigo Vergara, Jennifer Tate, Michelle McMahon, and Kevin Smith for providing fundamental plant material for this study. I wish to thank my colleagues of the Universidad de la República, Uruguay, particularly Magdalena Vaio, Cristina Mazzella, and Iván Grela for facilitating plant material and staying constantly in contact with me. This work would not have been possible without the training and inspiration that I received from Juan Carlos Millot during my undergraduate education. Finally, this dissertation was possible due to the support I received from my wife, Alicia Lusiardo. This work was supported in part by a credit-scholarship from the Programa de Desarrollo Tecnológico, Ministerio de Educación y Cultura, Uruguay. iv TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................................................................................. iv LIST OF TABLES........................................................................................................... viii LIST OF FIGURES ........................................................................................................... ix ABSTRACT....................................................................................................................... xi CHAPTER 1 AN INTRODUCTION TO THE STUDY OF THE DILATATA GROUP OF Paspalum ......................................................................................................................1 A Historical Perspective ...............................................................................................1 The Contributions of This Dissertation ........................................................................7 Chapter 2: The Technique .....................................................................................7 Chapter 3: The Germplasm ...................................................................................8 Chapter 4: The Complex .......................................................................................8 Chapter 5: The All-Important Pentaploid(s)..........................................................9 2 NUCLEAR AND CYTOPLASMIC MICROSATELLITE MARKERS FOR THE SPECIES OF THE DILATATA GROUP OF Paspalum (POACEAE).....................10 Introduction.................................................................................................................10 Materials and Methods ...............................................................................................13 Microsatellite Capture .........................................................................................13 Chloroplast Markers ............................................................................................14 Plant Material ......................................................................................................15 Amplification and Scoring ..................................................................................17 Results.........................................................................................................................18 Capture and Amplification Success.....................................................................18 Variability............................................................................................................24 Chloroplast Variability ........................................................................................25 Discussion...................................................................................................................26 Capture Efficiency...............................................................................................26 Amplification Profile...........................................................................................26 Non-Focal Loci....................................................................................................27 Nuclear Loci ........................................................................................................28 v Chloroplast Microsatellites..................................................................................30 Conclusions.................................................................................................................30 3 BREEDING SYSTEM AND POPULATION GENETIC STRUCTURE OF Paspalum dilatatum ssp. flavescens (POACEAE). ....................................................32 Introduction.................................................................................................................32 Materials and Methods. ..............................................................................................34 Collection Strategy ..............................................................................................34 Microsatellite Amplification ...............................................................................35 Data Analysis.......................................................................................................36 Results.........................................................................................................................37 Discussion...................................................................................................................43 Mating System.....................................................................................................43 Genetic Differentiation and Geographical Structure...........................................44 4 EVOLUTIONARY RELATIONSHIPS AND MECHANISMS IN THE DILATATA GROUP (Paspalum, POACEAE) .........................................................49 Introduction.................................................................................................................49 Materials and Methods ...............................................................................................53 Plant Material ......................................................................................................53 DNA Extraction and Microsatellite Analysis......................................................54 Data Analysis.......................................................................................................55 Results.........................................................................................................................56 Variability in the Tetraploids...............................................................................56 Variability in the Apomicts .................................................................................60 Relationships among Apomicts...........................................................................61 Heterozygosity of the Apomicts..........................................................................63 Discussion...................................................................................................................64 Evolutionary relationships among the sexual tetraploid biotypes.......................64 Genetic structure of the sexual tetraploids ..........................................................66 Variability within the apomicts ...........................................................................67 The addition of the X genome, apomixis, and the origin of pentaploid P. dilatatum ..........................................................................................................67 Paspalum dilatatum Uruguaiana and 59B...........................................................68 Paspalum dilatatum clone 2 ................................................................................70 Paspalum dilatatum Chirú...................................................................................70 Paspalum pauciciliatum ......................................................................................71 Paspalum dilatatum Torres .................................................................................72 Conclusions.................................................................................................................72 5 PENTAPLOID X TETRAPLOID HYBRIDIZATION CYCLES IN Paspalum dilatatum (POACEAE): EXPLAINING THE CURRENT AND FUTURE EVOLUTIONARY SUCCESS OF AN IMBALANCED POLYPLOID...................74 Introduction.................................................................................................................74 vi Materials and Methods ...............................................................................................76
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