Comparative analyses on the phenology, electrophysiology, and chemistry of the beech leaf-mining weevil (Orchestes fagi L.) and its novel host American beech (Fagus grandifolia Ehrh.) by Simon P. Pawlowski Thesis submitted in partial fulfillment of the requirements for the Degree of Master of Science (Biology) Acadia University Fall Graduation 2017 © by Simon P. Pawlowski, 2017 This thesis by Simon P. Pawlowski was defended successfully in an oral examination on 12 May 2017. The examining committee for the thesis was: ________________________________ Dr. Bobby Ellis, Chair ________________________________ Dr. Suzanne Blatt, External Reader ________________________________ Dr. Glenys Gibson, Internal Reader ________________________________ Dr. Kirk Hillier, Supervisor ________________________________ Dr. Jon Sweeney, Supervisor ________________________________ Dr. Michael Stokesbury, Acting Head This thesis is accepted in its present form by the Division of Research and Graduate Studies as satisfying the thesis requirements for the degree of Master of Science (Biology). ii I, Simon P. Pawlowski, grant permission to the University Librarian at Acadia University to reproduce, loan, or distribute copies of my thesis in microform, paper, or electronic formats on a non-profit basis. I, however, retain the copyright in my thesis. ____________________________ Author ____________________________ Supervisor ____________________________ Date iii Table of Contents List of tables ...................................................................................................................... vii List of figures ..................................................................................................................... ix Abstract ............................................................................................................................. xii List of abbreviations ........................................................................................................ xiii Acknowledgements ........................................................................................................... xv CHAPTER 1.0 INTRODUCTION ..................................................................................... 1 1.1 Background information ............................................................................................... 1 1.1.1 Beech leaf-mining weevil (Orchestes fagi L.) ................................................... 1 1.1.2 American beech (Fagus grandifolia Ehrh.) ....................................................... 4 1.2 Significance................................................................................................................... 6 1.3 Figures and tables ......................................................................................................... 8 1.4 References ................................................................................................................... 14 CHAPTER 2.0 DOES ORCHESTES FAGI HAVE A PIONEER SEX?.......................... 18 2.1 Introduction ................................................................................................................. 18 2.2 Materials and methods ................................................................................................ 20 2.2.1 Field trapping bioassays ................................................................................... 20 2.2.2 Sex ratios within an Orchestes fagi population and the validation of a non- invasive sexing technique ......................................................................................... 21 2.2.3 Y-tube olfactometer bioassays ......................................................................... 22 2.2.4 Statistical analyses ........................................................................................... 23 2.3 Results ......................................................................................................................... 23 2.3.1 Field trapping bioassays ................................................................................... 23 2.3.2 Sex ratios within an Orchestes fagi population ............................................... 24 2.3.3 Y-tube olfactometer bioassays ......................................................................... 25 2.4 Discussion ................................................................................................................... 25 2.5 Figures and tables ....................................................................................................... 30 2.6 References ................................................................................................................... 35 CHAPTER 3.0 BEHAVIOURAL BIOASSAYS ............................................................. 39 3.1 Introduction ................................................................................................................. 39 3.2 Materials and methods ................................................................................................ 43 3.2.1 Animals ............................................................................................................ 43 iv 3.2.2 Colour-tube bioassays ...................................................................................... 43 3.2.3 Y-tube olfactometer bioassays ......................................................................... 44 3.2.4 Field trapping bioassays ................................................................................... 45 3.2.5 Statistical analyses ........................................................................................... 47 3.3 Results ......................................................................................................................... 48 3.3.1 Colour-tube bioassays ...................................................................................... 48 3.3.2 Y-tube olfactometer bioassays ......................................................................... 48 3.3.3 Field trapping bioassays ................................................................................... 49 3.4 Discussion ................................................................................................................... 50 3.5 Figures and Tables ...................................................................................................... 57 3.6 References ................................................................................................................... 63 CHAPTER 4.0 A NOTE ON A NOVEL ELECTROANTENNOGRAM METHOD FOR ORCHESTES FAGI L. ...................................................................................................... 70 4.1 Introduction ................................................................................................................. 70 4.2 Materials and methods ................................................................................................ 71 4.3 Figures and tables ....................................................................................................... 74 4.3 References ................................................................................................................... 77 CHAPTER 5.0 CHEMICAL ANALYSES AND ELECTROPHYSIOLOGY ................ 81 5.1 Introduction ................................................................................................................. 81 5.2 Methods....................................................................................................................... 82 5.2.1 Volatile collection from Fagus grandifolia Ehrh. ........................................... 82 5.2.2 Animals ............................................................................................................ 84 5.2.3 Electrophysiological responses to F. grandifolia Ehrh. volatiles .................... 84 5.2.4 Chemical isolation and identification .............................................................. 85 5.2.5 Data analyses ................................................................................................... 85 5.3 Results ......................................................................................................................... 86 5.3.1 Volatile profiles of Fagus grandifolia Ehrh. at five developmental stages ..... 86 5.3.2 Electrophysiological responses to Fagus grandifolia Ehrh. volatiles ............. 87 5.3.3 Presence of unknown compounds .................................................................... 88 5.4 Discussion ................................................................................................................... 88 5.4.1 Volatile profiles of Fagus grandifolia Ehrh. at five developmental stages ..... 88 5.4.2 Electrophysiological responses to Fagus grandifolia volatiles ....................... 90 v 5.4.3 Presence of unknown compounds .................................................................... 93 5.5 Figures and Tables ...................................................................................................... 94 5.6 References ................................................................................................................. 100 CHAPTER 6.0 CONCLUSIONS ................................................................................... 104 6.1 Summary ................................................................................................................... 104 6.2 Future directions ......................................................................................................