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Waitea and Rhizoctonia Pathogens of Seashore Paspalum

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Waitea and Rhizoctonia Pathogens of Seashore Paspalum WAITEA AND RHIZOCTONIA PATHOGENS OF SEASHORE PASPALUM – THE ROLE OF SALINITY IN DISEASE EXPRESSION AND CHARACTERIZATION OF A NEW WAITEA CIRCINATA VARIETY CAUSING BASAL LEAF BLIGHT By STEVEN JOSEPH KAMMERER 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 Steven Joseph Kammerer 2 To my wife and best friend Rosemary, my greatest source of support and encouragement and to God for all his blessings and guidance in my life 3 ACKNOWLEDGMENTS I would like to thank my advisor, Phil Harmon for all his guidance and help but more importantly for accepting me as a graduate student. I also want to thank all the members of my graduate committee, Lee Burpee, David Norman, and Kevin Kenworthy for their help and support throughout this project. I also want to thank my fellow graduate students. Todd Cooper has been a great friend, and I thank him for all his advice and help maintaining the turfgrass for these studies. Norma Flor was also a good friend and lent me assistance. Other people who were helpful to me in this endeavor, Patti Rayside, Carol Stiles, Brenda Rutherford, and the superintendents at the golf courses I worked with at Old Palm, Boca West, Vero Beach Country Blub, Parkland, Tuscany Reserve, the Plantation at Somerset, Hammock Bay and the Oaks Club, for allowing me to sample and collect data over two years. Thanks to the University of Florida and to my employer Syngenta for seeing potential in me and the knowledge I would gain upon completion to justify funding my tuition for this endeavor. Lastly, I express my deepest appreciation to my wife, without her urging over the years, I would have never even attempted to return to school for such a daunting undertaking after being out of school for so long. Her and my children‟s patience, understanding and confidence in me for all the days and nights when I was gone, in addition to my already busy schedule as a full-time Syngenta employee, is a lot for any family to endure. During the lonely, frustrating nights away from home alone in a camper, their love, encouragement and support kept me going. 4 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 7 LIST OF FIGURES .......................................................................................................... 9 LIST OF ABBREVIATIONS ........................................................................................... 11 ABSTRACT ................................................................................................................... 12 CHAPTER 1 REVIEW OF LITERATURE .................................................................................... 14 Seashore Paspalum ............................................................................................... 14 Rhizoctonia ............................................................................................................. 18 Rhizoctonia and Turfgrass ...................................................................................... 23 Rhizoctonia solani ............................................................................................ 23 Rhizoctonia zeae and Rhizoctonia oryzae ....................................................... 27 Salinity Issues ......................................................................................................... 30 2 SURVEY FOR RHIZOCTONIA SPECIES ISOLATED FROM SEASHORE PASPALUM GOLF COURSES IN FLORIDA .......................................................... 35 Introduction ............................................................................................................. 35 Materials and Methods............................................................................................ 38 Turfgrass Sampling .......................................................................................... 38 Temperature, soil pH and EC determination .............................................. 39 Fungal isolations ........................................................................................ 40 Morphological Characterization and Groupings ................................................ 40 Molecular DNA methods and isolate identification ..................................... 41 Correlation of Edaphic Factors and Isolation Frequency of Fungi .................... 43 Results .................................................................................................................... 44 Isolates Recovered – Phylogenetics, Morphology and Descriptions ................ 44 Means and Isolate Recovery Frequencies ....................................................... 46 Isolate Recovery Correlations .......................................................................... 47 Discussion .............................................................................................................. 47 3 IDENTIFICATION OF A NEW WAITEA CIRCINATA VARIETY CAUSING BASAL LEAF BLIGHT OF SEASHORE PASPALUM ............................................. 63 Introduction ............................................................................................................. 63 Materials and Methods............................................................................................ 65 Turfgrass Sampling and Isolate Maintenance. ................................................. 65 5 Colony and Fungal Characteristics. .................................................................. 66 Internal Transcribed Spacer Region DNA Sequencing. ................................... 66 DNA Cloning. .................................................................................................... 67 Temperature, Growth Studies. ......................................................................... 68 Pathogenicity Studies. ...................................................................................... 68 Results .................................................................................................................... 71 Colony Morphology and Characterization......................................................... 71 rDNA-ITS Sequence Analysis. ......................................................................... 71 Turfgrass Pathogenicity Studies. ...................................................................... 72 Discussion .............................................................................................................. 73 4 THE INTERACTION OF CHRYSORHIZA ZEAE, RHIZOCTONIA SOLANI AG 2-2LP AND SALINE WATER ON PASPALUM VAGINATUM ................................. 85 Introduction ............................................................................................................. 85 Materials and Methods............................................................................................ 86 Pathogen Isolations and Identification .............................................................. 86 Inoculum Preparation ....................................................................................... 87 Turfgrass Establishment and Maintenance ...................................................... 88 Salt Treatments ................................................................................................ 88 Chrysorhiza zeae inoculations ................................................................... 89 Rhizoctonia solani AG 2-2LP inoculations ................................................. 90 Results .................................................................................................................... 91 Chrysorhiza zeae Salinity Experiment 1 ........................................................... 91 Chrysorhiza zeae Salinity Experiment 2 ........................................................... 92 Rhizoctonia solani AG 2-2LP Salinity Experiment 1 ......................................... 92 Rhizoctonia solani AG 2-2LP Salinity Experiment 2 ......................................... 93 Discussion .............................................................................................................. 94 LIST OF REFERENCES ............................................................................................. 104 BIOGRAPHICAL SKETCH .......................................................................................... 115 6 LIST OF TABLES Table page 1-1 “Rhizoctonia” taxonomy and nomenclature for fungal isolates in this dissertation ......................................................................................................... 34 2-1 Species utilized in this study and identification from sampling effort (or otherwise noted) utilizing rDNA internal transcribed spacer (ITS) region sequences for genetic characterization .............................................................. 51 2-2 Isolate recovery frequencies and SP tissue location .......................................... 52 2-3 Mean of Rhizoctonia-like fungus isolation recovered from seashore paspalum tissue, soil EC (salinity), soil pH, soil temperature, and canopy temperatures ... 52 2-4 Mean Rhizoctonia-like isolate recovery from golf course sites, EC and pH over nine sampling events .................................................................................
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