Characterising and understanding factors that affect the rate of molecular evolution in proteins has played a major part in the development of evolutionary theory. The early analyses of amino acid substitutions stimulated the development of the neutral theory of molecular evolution, which later evolved into the nearly neutral theory. More recent work has lead to a better understanding of the role selection plays at the molecular level, but there is still limited understanding of how higher levels of protein organisation affect the way natural selection acts. The investigation of this question is the central aim of this thesis, which is addressed via the analysis of selective pressures in secondary protein structures in insects, mammals and fungi. The analyses for the first two groups were conducted using publically available datasets. To conduct the analyses in fungi, genome sequence data from the fungal genus Microbotryum (sequenced in our laboratory) was assembled and annotated, resulting in the development of a number of bioinformatics tools which are described here. The fungal, insect and mammalian datasets were interrogated with regard to a number of structural features, such as protein secondary structure, position of a site with regard to adaptively evolving sites, hydropathy and solvent-accessibility. These features were correlated with the signals of positive and purifying selection detected using phylogenetic maximum likelihood and Bayesian approaches. I conclude that all of the factors examined can have an effect on the rate of molecular evolution. In particular, disordered and hydrophilic regions of the protein are found to experience fewer physiochemical constraints and contain a higher proportion of adaptively evolving sites. It is also revealed that positively selected residues are ‘clustered’ together spatially, and these trends persist in the three taxa. Finally, I show that this variation in adaptive evolution is a result of both selective events and physiochemical constraint.

In this thesis I discuss one way in which evolutionary theory has been brought to bear on the evaluation of competing meta-normative views in epistemology. More specifically, I investigate whether normative realism in epistemology (epistemic realism) is compatible with the view that we are justified in holding many of the epistemic beliefs we hold, on the assumption that those beliefs can be explained in evolutionary terms. In Part I I discuss normativity in epistemology and meta-epistemology. I begin by drawing attention to the fact that there are very different ways of understanding which concepts in epistemology are normative and what their normativity consists in. I focus on the concept of an epistemic reason to illustrate this point. I then discuss, in some detail, how different interpretations of epistemic-normativity will affect the form which normative-realist views in epistemology can take. I conclude by drawing a taxonomy of epistemic-realist views which is mindful of the different interpretations of epistemic normativity which I distinguished. In Part II I turn to discuss the topic of Darwinian arguments against normative-realist views in epistemology. I begin by considering the form which Darwinian arguments have taken in recent metaethical literature. I argue that Darwinian arguments of a kind which is meant to raise a distinctively epistemological challenge for normative-realist views – I call those 'Darwinian-epistemological' arguments – represent a more interesting object of philosophical investigation than Darwinian arguments of a different 'metaphysical' kind. I then formulate a Darwinian-epistemological argument which targets normative-realist views in epistemology (DEA), explain how it works, and spend some time discussing its key premises. In Part III I engage with the Darwinian-epistemological challenge against realism about epistemic normativity that Sharon Street presents in her paper 'Evolution and the Normativity of Epistemic Reasons' (2009). I argue that Street’s epistemological challenge is best viewed as an instance of a Darwinian-epistemological argument of the kind I formulated (DEA). I then go on to reply to Street’s argument on behalf of the epistemic realist. I conclude that arguments with the form of DEA do not represent a serious threat for normative realist views in epistemology.

Tropical forests have existed for up to one hundred million years, and today provide many ecosystem services vital for human well-being. They also harbour great biodiversity, which, in addition to its intrinsic value, plays a key role in the functioning of these ecosystems. Despite their local to global significance, there are still many knowledge gaps concerning the dynamic processes that govern the functioning of tropical forests. Rapid rates of deforestation and landscape conversion, predominantly for logging and industrial agriculture, are limiting the time and opportunity available to collect the information needed to fill these gaps. This research aims to shed light on the long-term ecological functioning of tropical forests, specifically investigating the history of disturbance in these ecosystems and the response of forest vegetation to past perturbations. The carbon-rich tropical peat swamp forests found along the coast of Sarawak, Malaysian Borneo, are a central focus of this study. For these forests in particular, a large deficit of knowledge surrounding their history and unique ecological functioning is coupled with some of the highest conversion rates of all tropical forest ecosystems across the world. In this thesis, palaeoecological data has been used to reconstruct temporal variability in forest vegetation coincident with external perturbations in order to identify changes in the resilience of these ecosystems through time, via indicators such as slowing rates of recovery and reduced regeneration of forest vegetation. Results suggest that tropical forest ecosystems have, for the most part, shown resilience to natural disturbances in the past, ranging from instantaneous localised tree-fall to longer-term regional climatic change; but that recent anthropogenic disturbances, of novel forms and greater intensities, are jeopardizing the potential for forest recovery and thus compromising ecosystem resilience. These findings enhance our understanding of the ecology of tropical peat swamp forests, and tropical forests more broadly. They also provide a context for contemporary tropical forest management, allowing for predictions on future responses to disturbance and enabling more ecologically sustainable landscape planning.

The allocation of resources to male or female progeny, or to male or female reproductive function more generally, is one of the most important life history decisions a sexually reproducing individual must ever make. Sex determination is thus a fundamental process, yet the mechanisms which control it are surprisingly diverse. In this thesis, I examine sex determination in the plant species Mercurialis annua L. (Euphorbiaceae). I assess the mechanism of sex determination operating in dioecious and androdioecious populations of M. annua and also investigate the conservation and evolution of sex-determining mechanisms across the annual mercury clade, the lineages of which display exceptional variation in sexual system. First, using crosses, I establish that sex in dioecious M. annua is controlled by a single-locus genetic mechanism, consistent with recent work that identified a single male-linked DNA marker in the species. My search for new sex-linked genes revealed none, however, suggesting that M. annua possesses at most a small non-recombining region around sex-determining loci. Why many dioecious plants lack heteromorphic sex chromosomes is still poorly understood and I consider explanations for this. I extend my investigation by comparing genetic diversity between loci that differ in their linkage to the sex-determining locus. I find a single male-linked marker to possess significantly lower diversity than autosomal loci, but no difference in the diversity of partially sex-linked and non-sex-linked genes. I also assess the conservation of a sex-linked marker among annual mercury lineages and conduct crosses between lineages to examine the conservation of sex determination. My findings indicate a conserved mechanism of single-locus genetic sex determination and I consider the role polyploidisation and hybridisation have played in sexual system evolution and the modification of sex-determining mechanisms in the clade. Finally, I assess the presence of environmental sex determination in androdioecious M. annua, concluding that although male frequency is not influenced by growing density, a degree of sexual lability exists in the lineage.

This dissertation clarifies the concept of evolvability, the increased capacity of some organisms or systems to support evolution, especially the evolution of life-like complexity. I survey the literature, which is spread over the fields of population genetics, developmental biology, artificial life, and microbial and molecular evolution. Finding that researchers have often used the term vaguely and incompatibly I identify five distinct kinds or senses of evolvability. I also identify five key constituent ideas, which I discuss in the context of organismic evolvability, a sense of evolvability with deep roots in the traditional fields of animal development and macroevolution. In these fields research into evolvability has historically been hampered by an insufficiently detailed knowledge of development. Research in molecular evolution has produced a thorough knowledge of the folding of RNA into secondary structure, which can be regarded as a model of development. This has motivated new approaches to evolvability based on representing development via a single genotype-phenotype mapping function. I build on these approaches to invent new mathematical methods to formalise the traditional ideas. I create an exact model illustrating a classic example of evolvability, the capacity for repeated segmentation and simple modularity. I analyse this with two new formal approaches. First is the genospace algebra, a propositional calculus based on graph theory. It is a formal language for describing genotype-phenotype maps. It provides a system for making calculations, proofs, and diagrams about mutational structures in genotype space, and it is flexible enough to allow description at arbitrary degrees of resolution. Second is a pair of concepts, the genetic leverage and the genetic fulcrum. The leverage provides a crude numerical measure of evolvability, and the fulcrum provides a heuristic for identifying the genomic and developmental causes of evolvability. Besides its specific relevance to diversification and development, evolvability is also crucial to the fundamental question of how evolution produces ordinary biological life. Simulation systems that implement only a conventional textbook model of evolution -– systems possessing only variation, inheritance, and selection –- fail to evolve anything resembling the complexity of the biological world. Research into evolvability is our best bet to illuminate the "missing ingredient" for life-like evolution.

In under 6 decades dengue has emerged from South East Asia to become the most widespread arbovirus affecting human populations. Recent dramatic increases in epidemic dengue fever have mainly been attributed to factors such as vector expansion and ongoing ecological, climate and socio-demographic changes. The failure to control the virus in endemic regions and prevent global spread of its mosquito vectors and genetic variants, underlines the urgency to reassess previous research methods, hypotheses and empirical observations. This thesis comprises a set of studies that integrate currently neglected and emerging epidemiological, ecological and evolutionary factors into unified mathematical frameworks, in order to better understand the contemporary population biology of the dengue virus. The observed epidemiological dynamics of dengue are believed to be driven by selective forces emerging from within-host cross-immune reactions during sequential, heterologous infections. However, this hypothesis is mainly supported by modelling approaches that presume all hosts to contribute equally and significantly to the selective effects of cross-immunity both in time and space. In the research presented in this thesis it is shown that the previously proposed effects of cross-immunological reactions are weakened in agent-based modelling approaches, which relax the common deterministic and homogeneous mixing assumptions in host-host and host-pathogen interactions. Crucially, it is shown that within these more detailed models, previously reported universal signatures of dengue's epidemiology and population genetics can be reproduced by demographic and natural stochastic processes alone. While this contrasts with the proposed role of cross-immunity, it presents demographic stochasticity as a parsimonious mechanism that integrates, for the first time, multi-scale features of dengue's population biology. The implications of this research are applicable to many other pathogens, involving challenging new ways of determining the underlying causes of the complex phylodynamics of antigenically diverse pathogens.

DNA methylation is an epigenetic modification that plays an important role in development and differentiation. The patterns of DNA methylation are largely established in early embryogenesis and maintained during development. Abnormal DNA methylation patterns have been associated with many human diseases, including cancer. Despite its importance, little is currently known about the mechanisms that determine DNA methylation patterns throughout the genome. To shed light on the molecular mechanisms that regulate DNA methylation, this study investigates whether DNA methylation patterns are established and maintained normally when human DNA is placed into a heterologous murine environment as opposed to its natural, endogenous chromosomal environment. Here, a previously generated transgenic mouse model, containing 117 kb of human DNA bearing the human α-globin cluster and all of its known regulatory elements, was analysed. The pattern of DNA methylation of the endogenous human α-globin cluster was compared with that of the transgenic cluster in the background of mouse embryonic stem cells (ESCs) and tissues. It was found that, although the normal human DNA methylation pattern was largely established and maintained in a mouse background, the region immediately around the human α-globin genes themselves is generally less methylated in mouse compared to human ESCs. It was found that regions adjacent and up to 2kb from the CpG islands (CGIs), so-called CGI shores, were unusually hypomethylated: this seems to be the result of an extension of CGIs in humanised mouse (hm) ESCs compared to human (h) ESCs. Furthermore, this hypomethylation appeared to increase during development in both erythroid and non-erythoid cells. To identify any cis-regulatory sequences responsible for the hypomethylated state of human CGI shores in the mouse, 2-4 kb human test sequences containing the CGI associated with the human α-globin 2 (α2) gene and its adjacent hypomethylated shore were re-integrated into the mouse α-globin locus via recombination-mediated cassette exchange (RMCE). Human CGI shores became hypomethylated in the context of the re-integrated test sequences, indicating that the appearance of hypomethylation is determined by the underlying human DNA sequence in the test fragments. In summary, the data presented here reveal that human CGIs become extended when placed in a mouse background leading to hypomethylation of human CGI shores in the mouse compared to the pattern of methylation at the normal endogenous human locus. These findings suggest that species-specific factors determine DNA methylation near CGIs. The transgenic mouse model provides an excellent system to dissect out species-specific regulation of CGI shore methylation. Furthermore, this study lays the foundation for future experiments addressing the role of DNA methylation in regulating human gene expression in the murine context, and examining the validity of transgenic mouse models for the study of human gene regulation.