Morphological and Molecular Characterisation of Australian Pinhole Borers (Coleoptera: Curculionidae, Platypodinae) James Ronald Munro Bickerstaff BSc, GradDipConBio Supervisors: A/Prof. Markus Riegler and Dr. Shannon Smith Thesis presented in partial fulfilment of the requirements for the degree of Master of Research April 10th 2017 Acknowledgements Taxonomic and systematic work is intricate, tiresome, and sometimes mind bending. One really needs a driving passion for the species that one is working on, otherwise, the cons outweigh the benefits of such elaborate work. If it were not for Shannon Smith I would never have gotten to know this fascinating group of insects exists. Not only this, you’ve been a fantastic supervisor. Thank you for your patience, for putting up with me constantly bursting into your office with countless questions, and showing me the ropes of molecular research. I look forward to continuing research with you on these enigmatic little beasties. And to Markus Riegler, thank you for your guidance, support, and tireless-ness. Your supervision and insights made this project what it is. Not only this, but your determination, energy, and belief in this project really helped give me the drive to continue on, especially through the rocky patches throughout the molecular lab work. Your advice you gave me during our meetings were amazingly helpful, and it was always entertaining when our meetings turned to discussions on global events and politics. I would like to also thank Jennifer Morrow; without your wisdom the molecular work in this project would not have happened. Your advice and help was greatly appreciated. To Robert Mueller, a PhD student in our little Platypodinae research group. Our discussions on the ecology and evolution of these beetles opened up many questions, some of which helped form the basis of this thesis. Additionally, thank you for letting me tag along on your fieldwork trips, even though there was no field work component to my research, it was good to learn how to collect pinhole borers in the field. To Ainsley Seago, for showing me the foundations of beetle morphology and illustration, and also for letting me go through the collections at Orange ASCU DPI to understand the diversity of Australian Pinhole borers. This project was reliant on utilising previously collected specimens stored in Natural History Collections. Without these fantastic collections the broad diversity of Australian Pinhole borers could not have been studied. Specifically, I would like to thank Ainsley Seago and Peter Gillespie (from Orange ASCU DPI), Derek Smith (from the Australian Museum), Justin Bartlett (from Queensland Primary Industries Insect Collection), Rolf Oberprieler and Debbie Jennings (from the ANIC CSIRO), and John Early and Dhahara Ranatunga (from the Aukland Museum) for organising borrowed specimens. I would also like to thank Geoff Monteith, who collected a great deal of Queensland material used in this study. And lastly, to Deborah Kent, who’s work on Austroplatypus incompertus has inspired research on Platypodinae in Australia; without your research, these cryptic weevils would have largely gone unnoticed. I would like to thank the Australian Biological Resources Study and Western Sydney University – Hawkesbury Institute of the Environment for project support. Statement of Authentication The work presented in this thesis is, to the best of my knowledge and belief, original except as acknowledged in the text. I hereby declare that I have not submitted this material, either in full or in part, for a degree at this or any other institution. James Bickerstaff 4 Table of Contents Table of Contents ..................................................................................................................... 4 List of Tables ............................................................................................................................ 7 List of Figures ........................................................................................................................... 9 Abbreviations ......................................................................................................................... 10 Preface ................................................................................................................................... 11 Thesis Abstract ....................................................................................................................... 13 Chapter 1: Introduction and background .............................................................................. 14 1.1 The evolutionary significance of weevils (Coleoptera: Curculionoidea) ...................... 14 1. 2. Who knows what weevils lurk in the hearts of trees – the natural history of Platypodinae (Coleoptera: Curcuilionidae, Platypodinae) ................................................. 17 1.2.1. Life history ............................................................................................................ 17 1.2.2. Fungiculture ......................................................................................................... 18 1.2.3. Behaviour and sociality ........................................................................................ 19 1.3. Ecological and economic significance of Platypodinae ............................................... 21 1.3.1 Tree pathogens transmitted by Platypodinae ...................................................... 22 1.3.2 Introduced and invasive Platypodinae .................................................................. 22 1.3.3. Management and prevention of pinhole borer attack ........................................ 23 1.4 Taxonomic, systematic, and evolutionary history of Platypodinae ............................. 24 1.4.1. Taxonomic history of Platypodinae ..................................................................... 24 1.4.2. Modern morphological and molecular phylogenetic analyses ............................ 25 1.4.3. Evolutionary history of Platypodinae ................................................................... 26 1.5 Species diversity in Australia ........................................................................................ 28 1.6 Rationale of this study ................................................................................................. 29 1.7 Study goals and chapter outlines ................................................................................. 31 1.7.1. Study goals ........................................................................................................... 31 Chapter 2: Host tree associations, distribution, and identification key of Australian Platypodinae .......................................................................................................................... 33 Abstract .............................................................................................................................. 33 2.1 Introduction ................................................................................................................. 33 2.2 Methods ....................................................................................................................... 35 2.2.1 Specimen acquisition ............................................................................................ 35 2.2.2 Compilation of host tree associations .................................................................. 36 2.2.3 Biogeography of pinhole borers ........................................................................... 36 2.2.4 Identification key development ............................................................................ 36 5 2.3 Results .......................................................................................................................... 37 2.3.1 Specimen availabilities .......................................................................................... 37 2.3.2 Host tree associations ........................................................................................... 38 2.3.3 Biogeography of Australian pinhole borers .......................................................... 38 2.3.4 Identification key of Australian pinhole borers .................................................... 43 2.4 Discussion ..................................................................................................................... 46 2.4.1 Specimen accessions in Natural History Collections ............................................. 46 2.4.2 Pinhole borer and host tree biogeography ........................................................... 47 2.4.3 Identification key of Australian pinhole borers .................................................... 51 Chapter 3: DNA barcoding and molecular systematics of Australian Platypodinae .............. 53 Abstract .............................................................................................................................. 53 3.1 Introduction ................................................................................................................. 53 3.2 Materials and Methods ................................................................................................ 57 3.2.1 Specimen collection and identification ................................................................. 57 3.2.2 DNA Extraction and amplification ......................................................................... 58 3.2.3 Sequence assembly and management