Cuscuta Pentagona Engelm.) USING GLYPHOSATE, AMMONIUM SULFATE, and the BIOLOGICAL CONTROL AGENT Alternaria Destruens Simmons, Sp
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INTEGRATED CONTROL OF DODDER (Cuscuta pentagona Engelm.) USING GLYPHOSATE, AMMONIUM SULFATE, AND THE BIOLOGICAL CONTROL AGENT Alternaria destruens Simmons, sp. nov. By JENNIFER COLLEEN COOK 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 2006 To my husband, Jeffrey Cook, for his love, understanding, and support. I could not have done this without you. ACKNOWLEDGMENTS I would like to sincerely thank Dr. Raghavan Charudattan for his guidance, patience, and support. In addition, I would like to express my deepest gratitude to Dr. Erin Rosskopf, without whose support I would never have made it on my own through this journey. I can’t thank her enough. I would also like to acknowledge the members of my committee, Drs. Greg MacDonald, William Stall, and Thomas Zimmerman for their support and guidance. I would like to thank Dr. Tom Bewick for his counsel and unending information on dodder; Jim DeValerio for his friendship and always being there when I had a problem or needed an answer, and for his experimental and statistical knowledge; Eldon Philman and Herman Brown for all of their assistance at the greenhouse, including building some crazy stuff for me; Philip Ruck for providing me citrus plants whenever I needed them; Camilla Yandoc for all her time in helping me with my field study; Sylvan and John Cascino for providing Smolder™ to accomplish this work; Dr. Portier for his statistical consulting services; Alana Den Breeyen, Abby Guerra, and Linley Smith for all their time and help in getting me through the molecular part of this research; Everyone in Charu’s lab, past and present, who has assisted me with this project; USDA-CSREES-Special Grants Program for the award of a TSTAR grant to the University of the Virgin Islands and the University of Florida that supported this research; and USDA-ARS-USHRL for a cooperative research agreement that partially supported this research. A special thanks to Whitney Elmore for her friendship, input, and shared misery of writing a dissertation. I might just miss those midnight emails! I iii would also like to thank my closest friends, Angela Horton, Lesa Werkmeister, Lynn Howard, Jill Bejarano, and Kim Rosiek for listening to me when I needed to cry, scream, or whine. Finally, I would like to thank my mom Catherine who has always been there no matter what, through good and bad times, and to push me to finish this degree when I needed the motivation. Mom, you’re my inspiration and I love you for it. iv TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................................................................................. iii LIST OF TABLES............................................................................................................ vii LIST OF FIGURES ........................................................................................................... ix ABSTRACT...................................................................................................................... xii CHAPTER 1 LITERATURE REVIEW .............................................................................................1 Taxonomy.....................................................................................................................1 Biology and Host Range...............................................................................................2 Cuscuta pentagona Engelm..........................................................................................6 Cuscuta spp. in the U. S................................................................................................7 Cuscuta spp. Worldwide...............................................................................................8 Economic Loss, Dispersal, and Control .......................................................................9 Herbicides...................................................................................................................10 Glyphosate ..................................................................................................................13 Ammonium Sulfate.....................................................................................................14 Alternaria destruens Simmons, sp. nov......................................................................15 Biological Control ......................................................................................................17 Research Objective .....................................................................................................18 2 EVALUATION OF THE HOST RANGE OF Alternaria destruens (SMOLDER™)............................................................................................................20 Materials and Methods ...............................................................................................23 Results and Discussion ...............................................................................................25 3 FIELD EFFICACY OF Alternaria destruens (SMOLDER™) AS A BIOHERBICIDE OF Cuscuta....................................................................................31 Materials and Methods ...............................................................................................32 Results and Discussion ...............................................................................................39 v 4 MOLECULAR PHYLOGENIC CHARACTERIZATION TO DIFFERENTIATE Alternaria destruens Simmons, sp. nov. FROM VARIOUS Alternaria SPECIES BASED ON INTERNAL TRANSCRIBED SPACER REGION ..............................50 Materials and Methods ...............................................................................................52 Results and Discussion ...............................................................................................56 5 DISPERSAL OF Alternaria destruens: A WIND TUNNEL STUDY......................65 Materials and Methods ...............................................................................................67 Results and Discussion ...............................................................................................69 6 EFFECTS OF Alternaria destruens (SMOLDER™), GLYPHOSATE (ROUNDUP PRO®), AND AMMONIUM SULFATE INDIVIDUALLY AND INTEGRATED FOR CONTROL OF Cuscuta pentagona...................................................................75 Materials and Methods ...............................................................................................80 Results and Discussion ...............................................................................................82 APPENDIX A FEDERAL NOXIOUS WEED LIST (AS OF JANUARY 6, 2006) ........................102 B WIND SPEED DATA...............................................................................................103 C ENVIRONMENTAL DATA FOR GREENHOUSE TRIALS OF Alternaria destruens, AMMONIUM SULFATE AT 0.125% W/V IN WATER, AND GLYPHOSATE AT 0.02 kg (ae)/L OF ROUNDUP PRO® ON Cuscuta pentagona .................................................................................................................107 D DATA FROM PRELIMINARY GREENHOUSE TRIAL ......................................111 LIST OF REFERENCES.................................................................................................119 BIOGRAPHICAL SKETCH ...........................................................................................130 vi LIST OF TABLES Table page 2-1. Reaction of test plants to Alternaria destruens applied as SMOLDER™ formulated with PCC588 (United Agri Products, Greeley, CO)..............................29 3-1. Means of area under the disease progress curve (AUDPC) for treatments on Cuscuta pentagona from Ft. Pierce field trial I........................................................45 3-2. Means of area under the disease progress curve (AUDPC) for treatments on Cuscuta pentagona from Ft. Pierce field trial II ......................................................46 4-1. Sources of unknown isolates and treatments applied to plots from which these isolates were recovered ............................................................................................60 4-2. Sources of Alternaria spp. used for DNA sequencing in this study..........................61 4-3. Fungal Isolates and the GenBank accession numbers for sequences used in phylogenic analyses..................................................................................................63 6-1. Regression equations and R2 values from greenhouse trials I and III (combined) for the effects of disease or damage severity from treatments on Cuscuta pentagona over time (days after treatment, DAT) ...................................................89 6-2. Area under the disease or damage progress curve (AUDPC) means for all treatments on Cuscuta pentagona from greenhouse trials I and III (combined)......91 6-3. Area under the disease or damage progress curve (AUDPC) means for all treatments on Cuscuta pentagona from greenhouse trial II .....................................93 B-1. Ft. Pierce trial I average wind speed data ...............................................................103 B-2. Ft. Pierce trial II average wind speed data..............................................................105 B-3. Gainesville field study average wind speed data ....................................................106 D-1. Area under the disease or