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Relationships and Biology of Ambrosia Beetles and Fungi and the Development of Pre-Invasion Assessment of Potential Pests

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RELATIONSHIPS AND BIOLOGY OF AMBROSIA BEETLES AND FUNGI AND THE DEVELOPMENT OF PRE-INVASION ASSESSMENT OF POTENTIAL PESTS By YOU LI 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 2018 © 2018 You Li To my wife and supportive parents ACKNOWLEDGMENTS There are so many people who have made this effort possible, and I would not have enough room to name them all. First, I must thank my wife, Misa, who stood by me during this four years. I do not know where I would be without her support and wisdom. My major advisor, Dr. Jiri Hulcr, has been a tremendous mentor and I feel fortunate he was kind enough to take me on as a student. His experience, and generosity throughout this process ensured the success of this. I would like to thank my committee members for their guidance and patience over the last few years. This includes Dr. Damian Adams, Dr. Huiping Yang, Dr. Jason A. Smith, and Dr. Matthew E. Smith. Every member of my committee was very supportive and helped me grow during my Ph.D. work. Thank you for your time, expertise, and wisdom. The samples of my research are most from China and USA. Many people help me in each chapter. In Chapter 2, I am grateful to D. Rabern Simmons (University of Michigan) for teaching me how to write a manuscript. I would like to thank Changlin Zhao and Baokai Cui (Beijing Forestry University, China) for sharing molecular data and Lukas Stelinski and Chris Gibbard provided several specimens. In Chapter 3, I gratefully acknowledge Jianjun Guo (Guizhou University, China), Ki-Jeong Hong and Moo-Sung Kim (Suncheon National University), Pham Hong Thai and Tran Thi Men (Vietnam National Museum of Nature) for their assistance in sample collecting and logistic support. Roger Beaver helped us identify Ambrosiophilus. In Chapter 4, I would like to thank Jian Yao and Menglei Zhang (Institute of Zoology, Chinese Academy of Sciences) for facilitating access to the collection records. 4 Additional thanks to Angie Macias and Matt Berger (West Virginia University) for help with DNA sequencing and molecular identification. In Chapter 5, I would like to thank Adam Black and M. Patrick Griffith for their help and permission collecting beetles at the Montgomery Botanical Center (Miami, FL). I thank the Pathology Laboratory for Translational Medicine at West Virginia University’s School of Medicine for their assistance in histological preparations. Additional thanks to Thomas H. Atkinson (University of Texas, USA) for training on platypodine identification. In Chapter 6, I am grateful to Guangyu Liu, Jianjun Guo (Guizhou University) and Paige Carlson (University of Florida) for aid in beetle collections. In Chapter 7, I gratefully acknowledge Chengxu Wu and Zhen Zhang (Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, China), Bo Wang (Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences), Zaihua Yang (Guizhou Academy of Forestry), Jianghua Sun (Institute of Zoology, Chinese Academy of Sciences), Chi-yu Chen and Hou-Feng Li (National Chung Hsing University), Liang-Jong Wang (Taiwan Forestry Research Institute) and Ching-Shan Lin for their assistance in sample collecting and logistic support. I am thankful for the continuous support from the members of Forest Entomology Laboratory, School of Forest Resources and Conservation, University of Florida, my colleagues, Allan Gonzalez, Andrew J. Johnson, Caroline Storer, Craig Bateman, Demian Gómez, James Skelton, Sedonia Steininger, Yin-Tse Huang, Zachary Nolen. I am grateful to each of them for their candid assistance at every stage of this project. 5 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 9 LIST OF FIGURES ........................................................................................................ 10 LIST OF ABBREVIATIONS ........................................................................................... 11 ABSTRACT ................................................................................................................... 12 CHAPTER 1 INTRODUCTION .................................................................................................... 14 2 SYMBIOTIC FUNGI ASSOCIATED WITH AMBROSIODMUS IN USA .................. 17 Introduction to Chapter 2 ........................................................................................ 17 Materials and Methods for Chapter 2 ...................................................................... 18 Isolation and Culturing of Fungi from Ambrosiodmus ....................................... 18 Identification of Fungi from Ambrosiodmus ...................................................... 19 Microtome and Micro-CT .................................................................................. 20 Morphology of the Fungi from Isolation ............................................................ 22 Results for Chapter 2 .............................................................................................. 22 Isolation and Culturing of Fungi from Ambrosiodmus and Plants in the USA ... 22 Phylogenetic Analyses of Fungi from Ambrosiodmus in the USA .................... 23 Examination of Mycangia of Ambrosiodmus ..................................................... 23 Morphology of Fungi from Ambrosiodmus ........................................................ 23 Discussion for Chapter 2 ......................................................................................... 24 3 SYMBIOTIC FUNGI ASSOCIATED WITH AMBROSIODMUS IN ASIA ................. 32 Introduction to Chapter 3 ........................................................................................ 32 Materials and Methods to Chapter 3 ....................................................................... 34 Results to Chapter 3 ............................................................................................... 35 Identification and Phylogenetic Analyses of Symbiotic Fungus Flavodon ambrosius in Asia .......................................................................................... 35 Biology of Fungus Flavodon ambrosius in Asia ................................................ 36 Discussions to Chapter 3 ........................................................................................ 36 4 DISTRIBUTION, HOST AND SYMBIOTIC FUNGI OF EUWALLACEA FORNICATUS IN CHINA ........................................................................................ 42 Introduction to Chapter 4 ........................................................................................ 42 6 Materials and Methods to Chapter 4 ....................................................................... 43 Results and Discussions to Chapter 4 .................................................................... 43 5 SYMBIOTIC FUNGI ASSOCIATED WITH PLATYPODINAE IN THE SOUTHEASTERN USA .......................................................................................... 49 Introduction to Chapter 5 ........................................................................................ 49 Materials and Methods to Chapter 5 ....................................................................... 51 Platypodine Beetle Collection ........................................................................... 51 Isolation, Identification, Taxa Assignment, and Phylogenetic Analyses of Fungi Associated with Platypodine Beetle .................................................... 52 Community Analysis of Fungi from Platypodine Beetle .................................... 54 Results to Chapter 5 ............................................................................................... 55 Discussion to Chapter 5 .......................................................................................... 56 6 DEVELOPMENT OF MYCANGIUM IN XYLOSANDRUS AMBROSIA BEETLES .. 64 Introduction to Chapter 6 ........................................................................................ 64 Materials and Methods to Chapter 6 ....................................................................... 65 Experiment 1: Morphological Variation of Mycangia Test ................................. 65 Experiment 2: Mycangial Development Test .................................................... 66 Hand Dissection of Xylosandrus ....................................................................... 67 Micro-CT Imaging ............................................................................................. 67 Identification of Fungi from Mycangia ............................................................... 68 Results to Chapter 6 ............................................................................................... 68 Morphological Variation of Xylosandrus Mycangia ........................................... 68 Mycangial Development and Fungal Identification ........................................... 69 Discussions to Chapter 6 ........................................................................................ 69 7 PATHOGENICITY EVALUATION OF CHINESE BARK AND AMBROSIA BEETLE-VECTORED FUNGI IN USA .................................................................... 79 Introduction to Chapter 7 .......................................................................................
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  • Effects of Sugars and Salt on the Production of Glycosphingolipids in Mariannaea Elegans

    Effects of Sugars and Salt on the Production of Glycosphingolipids in Mariannaea Elegans

    Trace Nutrients Research 31 : 37−44（2014） Original Article Effects of Sugars and Salt on the Production of Glycosphingolipids in Mariannaea Elegans 1, 2） 1） 1, 2） 1, †） Yasushi Tani , Yasunori Yamashita , Shigeki Saito and Hisaaki Mihara 1）Department of Biotechnology, College of Life Sciences, Ritsumeikan University ＊ , 2）Ritsumeikan Global Innovation Research Organization, Ritsumeikan University ＊ Summary Glycosphingolipids (GSLs) are complex macromolecules in cell membranes that play important roles in various bio- logical processes. Fungi contain several types of GSLs that are distinct from those of mammals, but little is known about their physiological functions, metabolic regulation, or biosynthetic pathways. Most fungal species that possess fungal neogala-series GSLs (FNG-GSLs) are resistant to aureobasidin A (AbA), an antifungal agent that inhibits glyco- sylinositolphosphoceramide (GIPC) synthesis. We have previously reported that Mariannaea elegans has FNG-GSLs in its cell membrane, despite the fact that it is sensitive to AbA. In this study, we demonstrated that M. elegans contains GIPCs, which explains its sensitivity to AbA. We also found that both cell growth and GSL composition of M. elegans were affected by the presence or absence of different sugars in the culture medium. In contrast, sugars had no effect on the phenotypes and GSLs of AbA-resistant Rhizopus sp. lacking GIPCs. Further analysis revealed that FNG-GSL production can be modulated by the addition of 600 mM NaCl in a glucose-containing medium. Fi- nally, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of Golgi proteins from M. elegans showed that at least nine proteins were present exclusively in cells producing FNG-GSL, suggesting that those proteins are potential candidates for FNG-GSL synthesis.