It is likely that the strength of selection acting upon a mutation varies through time due to changes in the environment. However, most population genetic theory assumes that the strength of selection remains constant. Here we investigate the consequences of fluctuating selection pressures on the quantification of adaptive evolution using McDonald-Kreitman (MK) style approaches. In agreement with previous work, we show that fluctuating selection can generate evidence of adaptive evolution even when the expected strength of selection on a mutation is zero. However, we also find that the mutations, which contribute to both polymorphism and divergence tend, on average, to be positively selected during their lifetime, under fluctuating selection models. This is because mutations that fluctuate, by chance, to positive selected values, tend to reach higher frequencies in the population than those that fluctuate towards negative values. Hence the evidence of positive adaptive evolution detected under a fluctuating selection model by MK type approaches is genuine since fixed mutations tend to be advantageous on average during their lifetime. Never-the-less we show that methods tend to underestimate the rate of adaptive evolution when selection fluctuates.

Mutations are the ultimate source of new genetic information and they can be neutral,
harmful or beneficial. The ultimate fate of all mutations is either to be lost or to eventually
become fixed in a population. In this thesis I investigate genome wide traces of natural
selection in eukaryotes. I focus on the most common type of mutations, point mutations,
in protein coding genes.

I investigated whether there is adaptive evolution in 11 plant species comparisons by
applying an extension of the McDonald Kreitman (MK) test and found little evidence
of adaptive evolution. However, most of the investigated plant species have low effective
population sizes (Ne) and the rate of adaptive evolution is thought to be correlated to
Ne. I therefore extended my study using additional data from mammals, drosophilids
and yeast to investigate the relationship between the rate of adaptive evolution and Ne.
I found a highly significant correlation between the rate of adaptive evolution relative to
the rate of neutral evolution (!a) and Ne.

It has been proposed that evidence of adaptive evolution can be an artifact of fluctuating
selection. I simulated a model of fluctuating selection, in which the average strength
of selection acting upon mutations is zero. Under this model adaptive evolution is inferred
using MK-type tests. However, the mutations which become fixed are on average positively
selected. The signal of adaptive evolution is therefore genuine.

Ne can not only vary between species but also across genomes. However, how much
variation there is, and whether this affects the efficiency of natural selection, is unknown.
I analysed 10 species and show that variation in Ne is widespread. However, this variation
is limited, amounting to a few fold variation in Ne between most genomic regions. This is
never-the-less sufficient to cause variation in the efficiency of selection.

The role of adaptation is a fundamental question in molecular evolution. Theory predicts that species with large effective population sizes should undergo a higher rate of adaptive evolution than species with low effective population sizes if adaptation is limited by the supply of mutations. Previous analyses have appeared to support this conjecture because estimates of the proportion of nonsynonymous substitutions fixed by adaptive evolution, α, tend to be higher in species with large N(e). However, α is a function of both the number of advantageous and effectively neutral substitutions, either of which might depend on N(e). Here, we investigate the relationship between N(e) and ω(a), the rate of adaptive evolution relative to the rate of neutral evolution, using nucleotide polymorphism and divergence data from 13 independent pairs of eukaryotic species. We find a highly significant positive correlation between ω(a) and N(e). We also find some evidence that the rate of adaptive evolution varies between groups of organisms for a given N(e). The correlation between ω(a) and N(e) does not appear to be an artifact of demographic change or selection on synonymous codon use. Our results suggest that adaptation is to some extent limited by the supply of mutations and that at least some adaptation depends on newly occurring mutations rather than on standing genetic variation. Finally, we show that the proportion of nearly neutral nonadaptive substitutions declines with increasing N(e). The low rate of adaptive evolution and the high proportion of effectively neutral substitution in species with small N(e) are expected to combine to make it difficult to detect adaptive molecular evolution in species with small N(e).

The effective population size (Ne) is one of the most fundamental parameters in population genetics. It is thought to vary across the genome as a consequence of differences in the rate of recombination and the density of selected sites due to the processes of genetic hitchhiking and background selection. Although it is known that there is intragenomic variation in the effective population size in some species, it is not known whether this is widespread or how much variation in the effective population size there is. Here, we test whether the effective population size varies across the genome, between protein-coding genes, in 10 eukaryotic species by considering whether there is significant variation in neutral diversity, taking into account differences in the mutation rate between loci by using the divergence between species. In most species we find significant evidence of variation. We investigate whether the variation in Ne is correlated to recombination rate and the density of selected sites in four species, for which these data are available. We find that Ne is positively correlated to recombination rate in one species, Drosophila melanogaster, and negatively correlated to a measure of the density of selected sites in two others, humans and Arabidopsis thaliana. However, much of the variation remains unexplained. We use a hierarchical Bayesian analysis to quantify the amount of variation in the effective population size and show that it is quite modest in all species—most genes have an Ne that is within a few fold of all other genes. Nonetheless we show that this modest variation in Ne is sufficient to cause significant differences in the efficiency of natural selection across the genome, by demonstrating that the ratio of the number of nonsynonymous to synonymous polymorphisms is significantly correlated to synonymous diversity and estimates of Ne, even taking into account the obvious nonindependence between these measures.

The relative contribution of advantageous and neutral mutations to the evolutionary process is a central problem in evolutionary biology. Current estimates suggest that whereas Drosophila, mice, and bacteria have undergone extensive adaptive evolution, hominids show little or no evidence of adaptive evolution in protein-coding sequences. This may be a consequence of differences in effective population size. To study the matter further, we have investigated whether plants show evidence of adaptive evolution using an extension of the McDonald-Kreitman test that explicitly models slightly deleterious mutations by estimating the distribution of fitness effects of new mutations. We apply this method to data from nine pairs of species. Altogether more than 2,400 loci with an average length of approximate to 280 nucleotides were analyzed. We observe very similar results in all species; we find little evidence of adaptive amino acid substitution in any comparison except sunflowers. This may be because many plant species have modest effective population sizes.