PHYSIOLOGICAL CHARACTERISTICS OF HERBICIDES AND MANAGEMENT OF OLD WORLD CLIMBING FERN (LYGODIUM MICROPHYLLUM) By JEFFREY T. HUTCHINSON 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 2010 1 © 2010 Jeffrey T. Hutchinson 2 To my wife, Thea, thanks for the love and support 3 ACKNOWLEDGMENTS First and foremost, I thank my wife, Thea, my most trusted friend, companion, supporter, and cosmic mate for 23+ years for her love and support. I am forever grateful and indebted to Dr. Kenneth Langeland for offering me the opportunity to pursue a Ph.D. in weed science. Dr. Langeland allowed me a unique opportunity to work independently, but always maintained an open door policy. My committee members all greatly contributed to the knowledge I learned over the last 5+ years. Dr. Robert Stamps took several days of his time to show me the basic culture techniques of fern propagation. Dr. Doria Gordon provided very insightful comments that I hope are exhibited throughout much of this dissertation. Dr. MacDonald offered great insight and comments on the design of experiments as well as allowing me to use his lab. Dr. William Haller offered insightful comments on different herbicides to screen. It is nearly impossible to recall all the assistance and support I received over the last 5+ years for field sites. Dr. Laura Brandt, Stefani Melvin, Gayle Martin, Dr. Mark Barrett, Jeremy Conrad, Lisa Jameson, Jon Wallace and all staff members at A.R.M. Loxahatchee N.W.R. provided the best support possible from 2005-2009. Kevin Maier, George Pelt and Don Napier were very helpful with the initial setup of plots on tree islands. Gayle Martin was a constant contact and went well beyond the call of duty many times when I needed help. I can’t think of a better place to do field research than A.R.M. Loxahatchee N.W.R. Dr. John Volin (now at the University of Connecticut) and his staff at Florida Atlantic University provided valuable insight on culture methods of OWCF spores. Dr. Michael Kane and his staff at the University of Florida provided valuable insight on 4 various methods that could be used to handle and count OWCF spores. Dr. Mete Yilmaz at the University of Florida gladly allowed me to use his lab on a moments notice, going well beyond the call of duty expected of him. Dr. Frank Chapman allowed me to use about every piece of equipment in his lab without ever making a minor grievance, and at the same time offering me very informative advice along the way. I am very thankful for the advice Dr. Chapman gave me. Bob Querns, David Mayo, Aimee Copper and Pedro Mendez at the University of Florida provided assistance in the lab and field, and I am grateful for their support. Bob Querns was very helpful with the translocation project and his help was greatly appreciated. Thea Hutchinson provided assistance in the field multiple times digging up OWCF plants in south Florida. Many staff and faculty members in the University of Florida Fisheries Department allowed me to use their equipment and provided practical advice, and I am grateful to all of them. I would like to thank the U.S. Fish and Wildlife Service, Florida Fish and Wildlife Conservation Commission’s Bureau of Invasive Plants, South Florida Water Management District, St. Johns River Water Management District, and the Florida Exotic Pest Plant Council for providing funding. Without the financial support of these agencies, these projects would not have been possible. Finally, I want to thank my parents, Melvia Hall and Cecil Hutchinson for their support over the years. My in-laws, now deceased, Helen and Amurat Bogushevitz were a constant source of moral and financial support, which made life easier. My crazy dogs, Maggie and Lola, made life fun and stress-free. I will always be grateful for the time I got to spend with Phillip Myers, a great friend and a big source of inspiration. 5 TABLE OF CONTENTS page ACKNOWLEDGMENTS.................................................................................................. 4 LIST OF TABLES............................................................................................................ 9 LIST OF FIGURES........................................................................................................ 12 ABSTRACT ................................................................................................................... 14 CHAPTER 1 INTRODUCTION .................................................................................................... 17 2 ABSORPTION AND TRANSLOCATION OF GLYPHOSATE, METSULFURON, AND TRICLOPYR IN OLD WORLD CLIMBING FERN (LYGODIUM MICROPHYLLUM).................................................................................................. 27 Introduction ............................................................................................................. 27 Materials and Methods............................................................................................ 30 Plant Material ................................................................................................... 30 Absorption and Translocation........................................................................... 31 Experimental Design and Data Analysis .......................................................... 32 Results and Discussion........................................................................................... 33 Visual Effects of Herbicides on OWCF............................................................. 33 Autoradiography ............................................................................................... 34 Absorption ........................................................................................................ 35 Translocation.................................................................................................... 36 3 TOLERANCE OF OLD WORLD CLIMBING FERN SPORES AND GAMETOPHYTES TO HERBICIDE EXPOSURE................................................... 46 Introduction ............................................................................................................. 46 Materials and Methods............................................................................................ 48 Source of Spores.............................................................................................. 48 Spore Preparation ............................................................................................ 49 Spore Counts ................................................................................................... 49 Germination Medium ........................................................................................ 49 Spore Inoculation ............................................................................................. 50 Germination Rates ........................................................................................... 51 Sporophyte Development................................................................................. 51 Experiment 1 - Tolerance of Gametophytes to Metsulfuron ............................. 52 Experiment 2 - Tolerance of Spores to Selected Herbicides ............................ 53 Experiment 3 - OWCF Spore Germination on Metsulfuron / Agar Medium ...... 54 Results.................................................................................................................... 55 Experiment 1 - Tolerance of Gametophytes to Metsulfuron ............................. 55 6 Experiment 2 - Tolerance of Spores to Selected Herbicides ............................ 56 Experiment 3 - OWCF Spore Germination on Metsulfuron / Agar Medium ...... 57 Discussion .............................................................................................................. 58 Experiment 1 - Tolerance of Gametophytes to Metsulfuron ............................. 58 Experiment 2 - Tolerance of Spores to Selected Herbicides ............................ 60 Experiment 3 - OWCF Spore Germination on Metsulfuron / Agar Medium ...... 62 4 REPEATED HERBICIDE APPLICATION FOR CONTROL OF OLD WORLD CLIMBING FERN AND THE EFFECTS ON NON-TARGET VEGETATION ON EVERGLADE’S TREE ISLANDS............................................................................ 83 Introduction ............................................................................................................. 83 Materials and Methods............................................................................................ 87 Study Site......................................................................................................... 87 Evaluation Periods ........................................................................................... 88 Herbicide Treatment......................................................................................... 88 Ground Cover................................................................................................... 89 Canopy Cover .................................................................................................. 89 Plant Species ................................................................................................... 90 Native Shrubs and Trees.................................................................................. 91 Statistical Analysis............................................................................................ 91 OWCF .............................................................................................................