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Dissertation.Pdf EFFECTS OF NATURAL/ANTHROPOGENIC STRESSORS AND A CHEMICAL CONTAMINANT ON PRE AND POST MYCORRHIZAL COLONIZATION IN WETLAND PLANTS Bishnu Ram Twanabasu, B.Sc., M.Sc., M.S Dissertation Prepared for the Degree of DOCTOR OF PHILOSOPHY UNIVERSITY OF NORTH TEXAS August 2013 APPROVED: Barney J. Venables, Major Professor Kevin J. Stevens, Major Professor Thomas La Point, Committee Member Jyoti Shah, Committee Member Duane Huggett, Committee Member Sam Atkinson, Chair of the Department of Biological Sciences Mark Wardell, Dean of the Toulouse Graduate School Twanabasu, Bishnu Ram. Effects of natural/anthropogenic stressors and a chemical contaminant on pre and post mycorrhizal colonization in wetland plants. Doctor of Philosophy (Biological Sciences – Plant Science), August 2013, 137 pp., 9 tables, 33 figures, references, 229 titles. Arbuscular mycorrhizal fungi, colonizing over 80% of all plants, were long thought absent in wetlands; however, recent studies have shown many wetland plants harbor arbuscular mycorrhizae (AM) and dark septate endophytes (DSE). Wetland services such as biodiversity, shoreline stabilization, water purification, flood control, etc. have been estimated to have a global value of $14.9 trillion. Recognition of these vital services is accompanied by growing concern for their vulnerability and continued loss, which has resulted in an increased need to understand wetland plant communities and mycorrhizal symbiosis. Factors regulating AM and DSE colonization need to be better understood to predict plant community response and ultimately wetland functioning when confronting natural and human induced stressors. This study focused on the effects of water quality, hydrology, sedimentation, and hurricanes on AM and DSE colonization in three wetland species (Taxodium distichum, Panicum hemitomon, and Typhal domingensis) and plant communities of coastal wetlands in Southeast Louisiana and effects of an antimicrobial biocide, triclosan (TCS), on AM (Glomus intraradices) spore germination, hyphal growth, hyphal branching, and colonization in fresh water wetland plants (Eclipta prostrata, Hibiscus laevis, and Sesbania herbacea) from bottom land hardwood forest in north central Texas. The former, mesocosm studies simulating coastal marsh vegetation ran for five years. In the latter studies, AM spores and wetland plants were exposed to 0 µg/L, 0.4 µg/L, and 4.0 µg/L TCS concentrations in static renewal and flow through exposures for 21 and 30 days, respectively. AM and DSE colonization was significantly affected by individual and interactions of four independent variables in mesocosm experiments. Similarly, spore germination, hyphal growth, hyphal branching, and AM colonization in selected wetland plants were significantly lowered by exposure to the TCS at environmentally relevant concentrations. However, levels of effects were plant species and fungal propagules specific. My results showed that natural and human induced alterations in environmental factors and chemical contaminants can significantly impact levels of mycorrhizal spore germination, colonization, and spore density in coastal and freshwater wetland plants. The resulting impacts on plant community structure and ecosystem function require further study. Copyright 2013 by Bishnu Ram Twanabasu ii ACKNOWLEDGEMENTS I would like to express my deep appreciation and gratitude to my major advisor, Dr. Kevin Stevens, for taking me on as a graduate student. His never ending advice, guidance, and encouragements during this study have directed me to explore my own path making this study a success. Furthermore, I cannot hold my emotional gratefulness to Dr. Steven’s family for great hospitality, accommodation, and food during my stay in Canada as part of this study. I would also like to gratefully and sincerely thank my second advisor Dr. Barney Venables for his continuous guardianship, assistance, and support especially after my major advisor took a job in Canada. I am also grateful to Dr. Jyoti Shah, Dr. Duane Huggett, and Dr. Thomas La Point for their outstanding advices and for serving on my dissertation committee. I thank Dr. Gary Shaffer, Bernard Wood, and Dr. Demetra Kandalepas for allowing me to collect samples from the mesocosm experiments and Mukesh, Ramjee, Sagar, and Prabesh to help collecting samples. Similarly, I am very grateful to UNT TAMS Keith, Joe, Bhargav, Trisha, Andrew, Eugene, and Amada for processing Louisiana samples and Alex and Susana preparing AM slides. I thank Caleb Smith and other lab mates for their great supports. I would like to thank Wilfrid Laurier University and UNT Biology to provide financial supports. I also thank Sanya Sidhu, Dr. William Sears, and Dr. Larry Peterson for their great contributions towards the manuscript preparation for publications. Finally, and most importantly, I’d be remiss if I didn’t acknowledge my parents Jaganath and Masinu, for their tremendous sacrifices that they made to ensure that I had an excellent education and brother Surya for his tremendous suggestions. I am indebted for great support, quiet patience, and unwavering love of my wife Geeta, dauthter Bishakha, and son Unik. iii TABLE OF CONTENTS Page ACKNOWLEDGEMENTS ................................................................................................................... iii LIST OF TABLES ............................................................................................................................... vii LIST OF FIGURES ............................................................................................................................ viii CHAPTER 1 INTRODUCTION ............................................................................................................ 1 1.1 Wetlands ................................................................................................................. 1 1.2 Mycorrhizas ............................................................................................................. 2 1.2.1 Arbuscular Mycorrhiza (AM) ....................................................................... 2 1.2.2 Colonization of Roots .................................................................................. 4 1.2.3 Dark Septate Endophytes (DSE) .................................................................. 5 1.3 Functions of Mycorrhizal Fungi ............................................................................... 6 1.3.1 Mycorrhizas in Wetlands ............................................................................ 7 1.4 Factors Affecting Mycorrhizal Colonization ............................................................ 9 1.5 Problem Statement and Objectives ...................................................................... 11 CHAPTER 2 EFFECTS OF WATER QUALITY, HYDROLOGY, SEDIMENTATION, AND A SIMULATED HURRICANE EXPOSURE ON ARBUSCULAR MYCORRHIZA (AM) AND DARK SEPTATE ENDOPHYTE (DSE) COLONIZATION IN COASTAL MARSH VEGETATION (Typha domingensis [Pers], Panicum hemitomon [L], Taxodium distichum [Schult]) .............................................................................. 14 2.1 Abstract ................................................................................................................. 14 2.2 Introduction .......................................................................................................... 15 2.3 Materials and Methods ......................................................................................... 19 2.3.1 Experimental Design ................................................................................. 19 2.3.2 Soil Sampling and Processing .................................................................... 22 2.3.3 Root Sampling and Processing .................................................................. 23 2.3.4 Plant Species Selection ............................................................................. 24 2.3.5 Data Analysis ............................................................................................. 25 2.4 Results ................................................................................................................... 25 2.5 Discussion.............................................................................................................. 37 2.6 Conclusions ........................................................................................................... 42 iv CHAPTER 3 TRICLOSAN INHIBITS ARBUSCULAR MYCORRHIZAL COLONIZATION IN THREE WETLAND PLANTS (Eclipta prostrata (L.) L., Sesbania herbacea (Mill.) Mcvaugh, AND Hibiscus laevis All) ....................................................................................................................................... 43 3.1 Abstract ................................................................................................................. 43 3.2 Introduction .......................................................................................................... 44 3.3 Methods and Materials ......................................................................................... 47 3.3.1 Plants ......................................................................................................... 47 3.3.2 Chemicals .................................................................................................. 48 3.3.3 Flow-Through Exposure System ............................................................... 48 3.3.4 Root Harvesting, Processing and AM Quantification ..............................
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