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This thesis is a comprehensive treatment of the invasion genetics of two major Liposcelis pest species, Liposcelis bostrychophila Badonnel and L. decolor (Pearman), in Australian grain storage systems. Randomly amplified polymorphic DNA (RAPDs) and microsatellite DNA markers were used to investigate Liposcelis invasions in grain storage systems. The RAPD and microsatellite markers used provided insights into the genetic diversity of L. bostrychophila and L. decolor populations both in Australia and internationally, providing information integral to gaining an understanding of Liposcelis invasions in Australian grain storage systems. The thesis is divided into discrete chapters, and for each chapter an abstract is provided. Chapter 1 provides background on Liposcelis invasions in Australia in relation to the biology of Liposcelis species, the infrastructure of the Australian grain industry and the history of invasions in comparison to other invasive invertebrate species. The use of DNA and PCR technologies to investigate Liposcelis invasions are discussed and the aims and objectives of this thesis are introduced. Chapter 2 uses RAPDs to trace the geographic origin of L. bostrychophila populations in Australia from unknown geographic sources internationally. High levels of clonal genetic diversity among populations of L. bostrychophila in Australia and internationally were found. In addition, multiple introductions, from a wide range of international source populations were detected and this obscured our ability to accurately determine the geographic origin of L. bostrychophila in Australia. Given the high clonal genetic diversity found in populations of parthenogenetic L. bostrychophila in Australia, diagnostic Wolbachia PCR primers were used in Chapter 3 to investigate whether L. bostrychophila individuals from these populations were infected by Wolbachia and if infected, to investigate the strain of Wolbachia characteristic of Australian L. bostrychophila populations. Results from Chapter 3 provide the first evidence of multiple Wolbachia infection from strains A and B in Australian L. bostrychophila populations. Chapter 4 details the extensive molecular procedures undertaken to isolate microsatellite loci from Liposcelis decolor using both enrichment and nonenrichment methods. Microsatellite loci were optimised for use in PCR in single individuals following extensive troubleshooting. Troubleshooting efforts focused on elucidating the factors controlling the specificity, efficiency and sensitivity of the PCR to amplify small Liposcelis individuals known to be rich in lipids and proteins, all inhibitory to PCR. In Chapter 5 lipids and proteins were investigated from L. decolor and L. entomophila to determine total concentrations and characterize the lipids from these species. This chapter discusses whether the lipid and protein concentrations found were of a level that could be inhibitory to PCR in relation to the microsatellite techniques used in this study. From the work conducted in both Chapters 4 and 5 a troubleshooting protocol adapted for use in L. decolor was developed and implemented to determine the endogenous and exogenous parameters responsible for the function and reproducibility of PCR of microsatellite loci in L. decolor. In Chapter 6, the novel microsatellites isolated from L. decolor in Chapter 4 were used to investigate genetic structure and gene flow from Australian and international L. decolor populations. In Chapter 6 the first evidence of population differentiation, gene flow and dispersal in invasive populations of L. decolor was found. In addition, the eleven microsatellites isolated from L. decolor were cross-amplified in five other important Liposcelis pests, L. bostrychophila, L. entomophila, L. paeta, L. rufa, and L. corrodens, from which informative population genetic studies are now possible. Finally, Chapter 7 comprises the thesis synopsis, implications and future research.

Selfish genetic elements can distort the sex ratios of their hosts by increasing their own transmission to the next generation in a non-mendelian fashion. These elements can be either nuclear genes on a sex chromosome or cytoplasmically inherited microbes, and achieve an increased transmission by manipulating gametogenesis or host reproduction. Often these selfish elements benefit from a female biased population (for example heritable microbes are passed on maternally in the egg cytoplasm), while non-selfish, autosomal genes are selected to produce a balanced sex ratio. These differing reproductive strategies cause a genetic conflict that results in an “evolutionary arms race” that can promote the evolutionary change of sex determination systems. In this thesis, I investigate an extreme sex ratio distortion in a species of booklouse, Liposcelis sp. This species contains two distinct female types, one of which carries a maternally transmitted selfish genetic element that results in exclusively female offspring being produced. Recently, a candidate for the sex ratio distortion was identified as a horizontally transferred bacterial gene, that we have called Odile, and that is present in the genome of the (distorter) female carrying the distorting element. The gene originates from the endosymbiotic bacterium Wolbachia that is well known for its ability to distort the sex ratio of its hosts. I investigated this horizontal gene transfer event and attempt to characterize Odile. I provide evidence that this Wolbachia gene has been integrated into the genome of the distorter females and is not a bacterial contaminant. I found that the Odile gene has been duplicated and may have been horizontally transferred from Wolbachia independently to at least three other insect genomes. Additionally, I found that Odile is transcribed at low levels in a life-stage specific manner that is suggestive of a role in development. Additionally, I looked into male mate choice in this species as one aspect of the persistence of the distorting element. I found that male Liposcelis sp. do not discriminate between the two female types and do not spend more time mating with one female type over the other. These results contribute to ongoing research into the extreme sex ratio distortion found in this species and the candidate gene that may be the cause. Selfish genetic elements are an important driver of sex determination evolution, and Liposcelis sp. provides a unique and exciting system to investigate the implications of selfish elements in a genome further. / Graduate / 2019-12-17