NEMATODES AFFECTING BANANA AND PLANTAIN AND MIRACLE FRUIT PRODUCTION IN SOUTH FLORIDA By LYNHE DEMESYEUX A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2019 © 2019 Lynhe Demesyeux To my loving mother for her endless love, sacrifices and for teaching me many valuable lessons that have guided me throughout my life ACKNOWLEDGMENTS I would like to thank the chair of my committee, Dr. Alan H. Chambers for being so patient and understanding towards me and helping me to achieve this goal. I also thank my committee members Dr. William T. Crow, Dr. Jonathan Crane and Dr. Randy Ploetz for their guidance and wisdom all along my time as a graduate student at UF. My friends Frantz Marc Penson Deroy, Cassandre Feuillé and Carina Theodore for their supports and the laughter shared. I also acknowledge Maria de Lourde Mendes PhD, Maria Brym and Sarah Brewer for their help in data collection, proof reading and support. Finally, I thank the AREA/Feed the future project for funding my research and Dr. Rosalie Koenig for her diligent administrative guidance. 4 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 8 LIST OF FIGURES .......................................................................................................... 9 LIST OF ABBREVIATIONS ........................................................................................... 10 ABSTRACT ................................................................................................................... 11 CHAPTER 1 LITERATURE REVIEW ........................................................................................... 13 Introduction to Banana and Plantain ....................................................................... 13 Introduction to Nematodes ...................................................................................... 14 Pathogenicity, Biology and Life Cycle of Plant Parasititc Nematodes on Banana and Plantain ..................................................................................... 16 Radopholus similis ........................................................................................... 16 Generalities ................................................................................................ 16 Symptoms and economic importance ........................................................ 17 Biology and life cycle ................................................................................. 17 Helicotylenchus spp. ........................................................................................ 18 Generalities ................................................................................................ 18 Symptoms .................................................................................................. 18 Biology and life cycle ................................................................................. 19 Generalities ................................................................................................ 19 Symptoms and economic importance ........................................................ 20 Biology and life cycle ................................................................................. 20 Meloidogyne spp. ............................................................................................. 20 Generalities ................................................................................................ 20 Symptoms and economic importance ........................................................ 21 Biology and life cycle ................................................................................. 21 Rotylenchulus reniformis .................................................................................. 21 Control Methods ............................................................................................... 21 Introduction to Miracle Fruit .................................................................................... 22 Introduction ....................................................................................................... 22 Botany .............................................................................................................. 23 Utilization .......................................................................................................... 24 Miraculin Extraction .......................................................................................... 24 Miraculin Purification ........................................................................................ 25 Potential Uses of Miraculin ............................................................................... 26 Taste-Altering Mechanism ................................................................................ 27 Current Medical Applications ............................................................................ 27 5 2 PLANT-PARASITIC NEMATODES ASSOCIATED WITH EIGHT BANANA CULTIVARS IN SOUTHERN FLORIDA.................................................................. 28 Summary ................................................................................................................ 28 Introduction ............................................................................................................. 29 Material and Methods ............................................................................................. 31 Site Selection ................................................................................................... 31 Musa Cultivars .................................................................................................. 32 Root Sampling .................................................................................................. 32 Nematode Extraction ........................................................................................ 32 Nematode Identification and Quantification ...................................................... 33 Preliminary Screening for Nematode Resistance ............................................. 33 Results .................................................................................................................... 34 Plant Parasitic Nematodes Nematodes from Soil Samples .............................. 34 Plant Parasitic Nematodes Associated with Musa Roots ................................. 35 Nematodes Associated with Musa Cultivars in Histosol and Limestone Soils .. 36 Preliminary Screening of Nematode Reproduction on Diverse Banana Accessions .................................................................................................... 37 Discussion .............................................................................................................. 44 Recognition ............................................................................................................. 47 3 YIELD PERFORMANCE AND MIRACULIN CONTENT OF A NOVEL FRUIT (SYNSEPALUM DULCIFICUM) IN HOMESTEAD, FLORIDA ................................ 48 Summary ................................................................................................................ 48 Introduction ............................................................................................................. 48 Material and Methods ............................................................................................. 50 Plant Material ................................................................................................... 50 Yield Evaluation ................................................................................................ 51 Miraculin Crude Extract .................................................................................... 51 Miraculin IMAC Purification and Protein Sequencing ....................................... 52 SDS-PAGE (Sodium Dodecyl Sulfate Polyacrilamide Gel Electrophoresis) ..... 52 Miraculin Quantification .................................................................................... 53 Statistical Analysis ............................................................................................ 53 Results .................................................................................................................... 53 Miracle Fruit Total Yield by Accession .............................................................. 53 Miracle Fruit Seasonal Yield ............................................................................. 54 Miracle Fruit Total Yield per Tree ..................................................................... 54 Miracle Fruit Average Fruit Weight ................................................................... 55 Protein Sequencing .......................................................................................... 55 Protein Extraction ............................................................................................. 55 Discussion .............................................................................................................. 59 Seasonal Yield ........................................................................................................ 59 Yield per Tree ..................................................................................................