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Trace Akankunda THE EVOLUTION OF DIET WIDTH IN LASIOGLOSSUM (CHILALICTUS) (HALICTIDAE, APOIDEA), IN ASSOCIATION WITH SPECIATION AND SENSORY MORPHOLOGY Trace Akankunda M.Bio (PB), The University of Adelaide Thesis submitted to The University of Adelaide for the degree of Doctor of Philosophy Department of Plant Protection School of Agriculture, Food & Wine Faculty of Sciences, The University of Adelaide Waite Campus April 2019 Table of contents List of tables ........................................................................................................................................................ 4 List of figures ...................................................................................................................................................... 5 List of supplementary material ................................................................................................................... 8 Tables ........................................................................................................................................................... 8 Figures ......................................................................................................................................................... 9 List of electronic files ................................................................................................................................... 10 Summary ........................................................................................................................................................... 13 Declaration ....................................................................................................................................................... 15 Acknowledgements ....................................................................................................................................... 17 Chapter 1: General Introduction - Why is broadly polylectic behaviour rare in bees? .... 21 Aims and overview ................................................................................................................................... 37 References ................................................................................................................................................... 39 Chapter 2: Multi-locus barcoding of pinned insect specimens using MiSeq ......................... 51 Abstract ......................................................................................................................................................... 52 Introduction ................................................................................................................................................ 53 Materials and Methods ........................................................................................................................... 58 Results ........................................................................................................................................................... 68 Discussion .................................................................................................................................................... 73 Acknowledgements .................................................................................................................................. 84 References ................................................................................................................................................... 85 Data Accessibility ...................................................................................................................................... 97 Appendices .................................................................................................................................................. 97 1 Supporting information ......................................................................................................................... 99 Chapter 3: The evolution of broadly polylectic behaviour in Lasioglossum (Chilalictus) (Halictidae, Apoidea)................................................................................................................................. 117 Abstract ...................................................................................................................................................... 118 Introduction ............................................................................................................................................. 119 Methods ..................................................................................................................................................... 123 Results ........................................................................................................................................................ 130 Discussion ................................................................................................................................................. 137 References ................................................................................................................................................ 143 Supporting information ...................................................................................................................... 151 Chapter 4: The association between broad polylecty and olfactory sensory morphology in Lasioglossum (Chilalictus) (Halictidae, Apoidea)...................................................................... 159 Abstract ...................................................................................................................................................... 160 Introduction ............................................................................................................................................. 161 Methods ..................................................................................................................................................... 165 Results ........................................................................................................................................................ 170 Discussion ................................................................................................................................................. 173 References ................................................................................................................................................ 176 Supporting information ...................................................................................................................... 182 CHAPTER 5 .................................................................................................................................................... 183 GENERAL DISCUSSION ............................................................................................................................. 183 Summary ................................................................................................................................................... 183 Why is broadly polylectic behaviour rare in bees? .................................................................. 184 2 Multilocus barcoding of pinned insect specimens using MiSeq ......................................... 185 The evolution of broad polylecty in Lasioglossum (Chilalictus) ......................................... 186 The association between pollen host breadth and olfactory sensory morphology in L. (Chilalictus) (Apoidea: Halictidae) .................................................................................................. 188 General Conclusion ............................................................................................................................... 189 References ................................................................................................................................................ 194 3 List of tables Table 1.1. Lexicon and criteria for classifying species according to their diet width (Cane & Sipes 2006). ............................................................................... 22 Table 2.1. Degenerate primers for Lasioglossum used to amplify CO1, EF-1α and WNT with references for those modified from published primers. Modifications include; adopting degenerate bases and adding M13 tails and Illumina sequencing adapters for next generation sequencing. For each gene region in the table, the first and last primers amplify near-full length loci and the middle primers are nested within each gene sequence to amplify 300-350 bp gene fragments. ....................................................... 60 Table 3.1. Species of Lasioglossum (Chilalictus) included in the sequence dataset for phylogenetic analysis with GenBank accession numbers. Sequences whose accession numbers start with MH were produced using the amplicon sequencing method developed in chapter 2, while the rest were sourced from GenBank archives. ........................................................................ 124 Table 3.2. A modified diet width classification system based on suggestions by Cane & Sipes (2006), that takes sampling bias into account. ......................... 127 Table 3.3. Summary of the categories of diet width for 77 Lasioglossum (Chilalictus) species. ................................................................................................... 133 Table 4.1. Phylogenetic signal of categorical and continuous traits using Pagel’s lambda.
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