Divergent patterns of diversification in courtship and genitalic characters of Timema walking‐sticks

Understanding the patterns of diversification in sexual traits and the selection underlying such diversification represents a major unresolved question in evolutionary biology. We examined the phylogenetic diversification for courtship and external genitalic characters across ten species of Timema walking‐sticks, to infer the tempos and modes of character change in these sexual traits and to draw inferences regarding the selective pressures underlying speciation and diversification in this clade. Rates of inferred change in male courtship behaviours were proportional to speciation events, but male external genitalic structures showed a pattern of continuous change across evolutionary time, with divergence proportional to branch lengths. These findings suggest that diversification of courtship behaviour is mediated by processes that occur in association with speciation, whereas diversification of genitalia occurs more or less continuously, most likely driven by forces of sexual selection.     

The evolution of host preference in allopatric vs. parapatric populations of Timema cristinae walking-sticks

Divergent habitat preferences can contribute to speciation, as has been observed for host-plant preferences in phytophagous insects. Geographic variation in host preference can provide insight into the causes of preference evolution. For example, selection against maladaptive host-switching occurs only when multiple hosts are available in the local environment and can result in greater divergence in regions with multiple vs. a single host. Conversely, costs of finding a suitable host can select for preference even in populations using a single host. Some populations of Timema cristinae occur in regions with only one host-plant species present (in allopatry, surrounded by unsuitable hosts) whereas others occur in regions with two host-plant species adjacent to one another (in parapatry). Here, we use host choice and reciprocal-rearing experiments to document genetic divergence in host preference among 33 populations of T. cristinae. Populations feeding on Ceanothus exhibited a stronger preference for Ceanothus than did populations feeding on Adenostoma. Both allopatric and parapatric pairs of populations using the different hosts exhibited divergent host preferences, but the degree of divergence tended to be greater between allopatric pairs. Thus, gene flow between parapatric populations apparently constrains divergence. Host preferences led to levels of premating isolation between populations using alternate hosts that were comparable in magnitude to previously documented premating isolation caused by natural and sexual selection against migrants between hosts. Our findings demonstrate how gene flow and different forms of selection interact to determine the magnitude of reproductive isolation observed in nature.     

Differential visual predation on morphs of Timema cristinae (Phasmatodeae:Timemidae) and its consequences for host range

Timema cristinae is a herbivorous insect that exhibits polymorphism for body coloration (green, red and grey morphs) and for pattern (striped, expressed only in the green morph, and unstriped). The striped green morph is associated with ceanothus ( Ceanothus spinosus ) and the unstriped green morph is associated with chamise ( Adenostoma fasciculatum ). This study examines the relative vulnerabilities to predation of the different pattern and colour morphs on their natural backgrounds. The vulnerabilities of the striped and unstriped morphs on their two food plants were tested using uncaged wild birds (Scrub Jays) and captive western fence lizards. Strong differential predation was observed suggesting that each morph is most cryptic on the food plant on which it is most common. Furthermore, in a mark-recapture experiment in a patch of ceanothus the unstriped and red morphs were recaptured in higher proportion than the other morphs. The vulnerabilities of the grey and green morphs on the ground and foliage were tested using lizards. The grey morph was more vulnerable on the plants than the green morph, but the inverse was observed on the ground (where they drop after a disturbance). This may be why the grey morph is not associated with specific food plants. The striped and colour polymorphisms in T. cristinae appear to be an evolutionary consequence of differential predation on different backgrounds. The implications of differential predation to food-plant utilization are discussed.     

Host choice promotes reproductive isolation between host races of the larch budmoth Zeiraphera diniana

The chances for sympatric speciation are improved if ecological divergence leads to assortative mating as a by-product. This effect is known in parasites that find mates using host cues, but studies of larch- and pine-feeding races of the larch budmoth (Zeiraphera diniana, Lepidoptera: Tortricidae) suggest it may also occur when mate attraction is via sex pheromones that are independent of habitat. We have previously shown that females releasing pheromones on or near their own host attract more males of their own race than if placed on the alternative host. This host effect would enhance assortative mating provided adults preferentially alight on their native hosts. Here we investigate alighting preferences in natural mixed forest using a novel likelihood analysis of genotypic clusters based on three semidiagnostic allozyme loci. Both larch and pine females show a realized alighting preference for their own host of 86%. The equivalent preferences of males were 79% for the larch race and 85% for the pine race. These preferences are also detectable in small-scale laboratory experiments, where alighting preferences of larch and pine races towards their own hosts were, respectively, 67 and 66% in females and 69 and 63% in males. Pure larch race moths reared in the laboratory had alighting choice similar to moths from natural populations, while hybrids were intermediate, showing that alighting preferences were heritable and approximately additive. The field estimates of alighting preference, coupled with earlier work on mate choice, yield an estimated rate of natural hybridization between sympatric host races of 2.2-3.8% per generation. Divergent alighting choice enhances pheromone-mediated assortative mating today, and is likely to have been an important cause of assortative mating during initial divergence in host use. Because resources are normally 'coarse-grained' in space and time, assortative mating due to ecological divergence may be a more important catalyst of sympatric speciation than generally realized.     

Lack of early laboratory postzygotic reproductive isolation between two ecotypes of Littorina saxatilis (Mollusca, Gastropoda) showing strong premating sexual isolation

Two sympatric and divergent adaptive ecotypes of Littorina saxatilis (RB and SU) are known to hybridize showing partial premating isolation in the wild. Previous studies have revealed that morphological intermediate forms (presumably hybrids) present fitness (viability, sexual selection and fecundity) similar to that from pure ecotypes at the mid-shore. However, the absence of postzygotic isolation due to genetic incompatibility cannot be ruled out unless it is measured directly on true F ₁ hybrids. In this study, we overcome this problem and present data on 56 individual crosses including the four possible mating combinations (RB/RB, RB/SU, SU/RB and SU/SU) to compare fertilization and fecundity rates (including young progeny viability) between the four type crosses. Pooled RB female crosses showed apparently larger fertility and fecundity than pooled SU female crosses, probably because of differences in fecundity and laboratory survivorship between ecotypes. However, similar fertilization and fecundity rates were found for both RB and SU females when mated with different male types, supporting the idea that genetic-incompatibility-based postzygotic isolation can be ignored as a major determinant of this polymorphism in nature.     

Divergent adaptation promotes reproductive isolation among experimental populations of the filamentous fungus <Emphasis Type=Italic>Neurospora</Emphasis>

Background  

An open, focal issue in evolutionary biology is how reproductive isolation and speciation are initiated; elucidation of mechanisms with empirical evidence has lagged behind theory. Under ecological speciation, reproductive isolation between populations is predicted to evolve incidentally as a by-product of adaptation to divergent environments. The increased genetic diversity associated with interspecific hybridization has also been theorized to promote the development of reproductive isolation among independent populations. Using the fungal model Neurospora, we founded experimental lineages from both intra- and interspecific crosses, and evolved them in one of two sub-optimal, selective environments. We then measured the influence that initial genetic diversity and the direction of selection (parallel versus divergent) had on the evolution of reproductive isolation.     

Divergent host plant specialization as the critical driving force in speciation between populations of a phytophagous ladybird beetle

Detecting the isolating barrier that arises earliest in speciation is critically important to understanding the mechanism of species formation. We tested isolating barriers between host races of a phytophagous ladybird beetle, Henosepilachna diekei (Coleoptera: Coccinellidae: Epilachnine), that occur sympatrically on distinct host plants. We conducted field surveys for the distribution of the beetles and host plants, rearing experiments to measure six potential isolating factors (adult host preference, adult and larval host performance, sexual isolation, egg hatchability, F(1) hybrid inviability, and sexual selection against F(1) hybrids), and molecular analyses of mitochondrial ND2 and the nuclear ITS2 sequences. We found significant genetic divergence between the host races, and extremely divergent host preference (i.e. habitat isolation) and host performance (i.e. immigrant inviability), but no other isolating barriers. The fidelity to particular host plants arises first and alone can prevent gene flow between differentiating populations of phytophagous specialists.     

Fluctuating asymmetry and reproductive isolation between two sticklebacks

This paper describes patterns of developmental asymmetry in a limnetic-benthic stickleback species pair from Paxton Lake, British Columbia. Three gill raker characters and one armor character were compared among full-sib parental crosses and their hybrids (F₁, F₂ and backcrosses). All crosses displayed asymmetry, but no differences in mean or fluctuating asymmetry were detected among laboratory-reared crosses. When wild-caught limnetics and benthics were included in the analyses significant differences in FA among groups were detected in gill raker length and plate number. The effect appears to be solely due to wild-caught benthics, though the reasons for their greater asymmetry are unknown. The patterns in laboratory-reared crosses suggest that asymmetry is unlikely to promote reproductive isolation between the parental species.     

The impact of flower-dwelling predators on host plant reproductive success

Flowers attract insects and so are commonly exploited as foraging sites by sit-and-wait predators. Such predators can be costly to their host plant by consuming pollinators. However, sit-and-wait predators are often prey generalists that also consume plant antagonists such as herbivores, nectar robbers and granivores, so may also provide benefits to their host plant. Here we present a simple, but general, model that provides novel predictions about how costs and benefits interact in different ecological circumstances. The model predicts that the ecological conditions in which flower-dwelling predators are found can generate either net benefits to their host plants, net costs to their host plants, or can have no effect on the fitness of their host plants. The net effect is influenced by the relative densities of mutualists and antagonists. The flower-dwelling predator has a strong positive effect on the plant if both the pollinators and the granivores are at high density. Further, the range of density combinations that yield a positive net outcome for the plant increases if the performance of pollinators is negatively density dependent, if the predator is only moderately effective at influencing flower visitor rates by its potential prey, and if pollinators are very effective. If plants of a given species find themselves consistently in conditions where they benefit from the presence of a predator then we predict that natural selection could favour the evolution of plant traits that increase the likelihood of predator recruitment and retention, especially where plants are served by highly effective pollinators.     

Choice of host plant as a factor in reproductive isolation of the aphid genus Cryptomyzus (Homoptera, Aphididae)

Abstract. 1. Host plant preference experiments were conducted with closely related taxa of the aphid genus Cryptomyzus. Males, and presexual morphs (sexuparae and gynoparae), were used to determine the impact of host plant choice on reproductive isolation. In the case of host-alternating species these morphs are migratory and so will select the host plant.
2. Host plant preference of two closely related taxa of C. alboapicalis (Theobald) was found to promote their reproductive isolation. The preference of sexuparae of these monoecious taxa was more pronounced than that of the males.
3. Host plant preference and subsequent production of oviparae showed that C. galeopsidis (Kaltenbach) consists of two host races restricted to Ribes rubrum L. and R. nigrum L., respectively. The existence of clones, intermediate in their preference and reproductive performance on these plants, suggests that hybridization occurs.     

Silicon deficiency and the adaptation of tropical rice ecotypes

Although silicon (Si) is found at much higher concentrations in healthy rice crops than N, P or K, it has received far less study, particularly for upland rice. There are few reports on the existence, causes, and effects of varying Si supplies in different environments. Chemical analyses of soil, water and plant tissue samples from experiments grown on a typical weathered, acidic upland soil in Colombia found concentrations of Si which were 80-90% lower than those in a typical lowland environment. These results corroborate published findings from West Africa and Hawaii, and lend support to a conclusion that acid-soil upland rice environments in the tropics tend to be deficient in Si, increasing disease damage, among other effects. Critical values for diagnosis of Si deficiency in soils, water and rice husk tissue are suggested. These are reasonably consistent with, but extend the application of previously published values derived from lowland rice studies to upland environments, and use simpler sampling and analysis methods. A strong correlation was found (r = -0.91) between high husk Si concentration and low husk discoloration disease damage, among diverse rice genotypes grown in the uplands. These genotypic differences were mainly explained by their ecotypic affinities: those belonging to the tropical japonica ecotype exhibited 93% higher husk Si concentrations than indica ecotypes (ecotypic means of 23 vs. 12 mg kg^-1 ). This is consistent with a hypothesis that the tropical japonicas may have adapted to Si deficiency in their native upland environment by evolving mechanisms to attain relatively higher tissue Si concentrations than indicas, which are believed to have evolved in the lowlands, where the Si supply is generally ample. Increased understanding of Si-mediated disease resistance in different rice environments and ecotypes could help breeders combine the high yield potential of indica types with the more durable disease resistance of the japonicas, and could contribute to the development of integrated disease management strategies.     

Evaluating the within-host fitness effects of mutations fixed during virus adaptation to different ecotypes of a new host

The existence of genetic variation for resistance in host populations is assumed to be essential to the spread of an emerging virus. Models predict that the rate of spread slows down with the increasing frequency and higher diversity of resistance alleles in the host population. We have been using the experimental pathosystem Arabidopsis thaliana—tobacco etch potyvirus (TEV) to explore the interplay between genetic variation in host''s susceptibility and virus diversity. We have recently shown that TEV populations evolving in A. thaliana ecotypes that differ in susceptibility to infection gained within-host fitness, virulence and infectivity in a manner compatible with a gene-for-gene model of host–parasite interactions: hard-to-infect ecotypes were infected by generalist viruses, whereas easy-to-infect ecotypes were infected by every virus. We characterized the genomes of the evolved viruses and found cases of host-driven convergent mutations. To gain further insights in the mechanistic basis of this gene-for-gene model, we have generated all viral mutations individually as well as in specific combinations and tested their within-host fitness effects across ecotypes. Most of these mutations were deleterious or neutral in their local ecotype and only a very reduced number had a host-specific beneficial effect. We conclude that most of the mutations fixed during the evolution experiment were so by drift or by selective sweeps along with the selected driver mutation. In addition, we evaluated the ruggedness of the underlying adaptive fitness landscape and found that mutational effects were mostly multiplicative, with few cases of significant epistasis.     

Proteomic comparison between two marine snail ecotypes reveals details about the biochemistry of adaptation

The proteomic changes occurring during speciation are fundamental to understand this process, though they have been rarely addressed until present. Therefore, we compared the proteome of two ecotypes (RB and SU) of the marine snail Littorina saxatilis, a case of sympatric incomplete speciation, originated as a byproduct of adaptation to distinct habitats. Thus, the RB ecotype is able to resist stresses of desiccation and temperature on the upper shore, whereas the SU ecotype defies strong physical disturbances due to wave action. Qualitative analyses of 2-DE gels demonstrated 21 proteins differentially expressed (1.4% of the proteome, 1.2% after considering type-I errors), while quantitative changes accounted for differences in 22 spots (16% of the proteome, 11% after considering type-I errors). These results suggest that adaptative phenotypic plasticity, natural selection, or both maintain these ecotypes in sympatry. Among the proteins identified by MS, we found that fructose-bisphosphate aldolase and arginine kinase were up-regulated in the SU ecotype, suggesting an enhancement of the level of energy available as ATP, in order to withstand its wave-exposed habitat.     

Metapopulation-level adaptation of insect host plant preference and extinction-colonization dynamics in heterogeneous landscapes

Species living in highly fragmented landscapes typically occur as metapopulations with frequent turnover of local populations. The turnover rate depends on population sizes and connectivities, but it may also depend on the phenotypic and genotypic composition of populations. The Glanville fritillary butterfly (Melitaea cinxia) in Finland uses two host plant species, which show variation in their relative abundances at two spatial scales: locally among individual habitat patches and regionally among networks of patches. Female butterflies in turn exhibit spatial variation in genetically determined host plant preference within and among patch networks. Emigration, immigration and establishment of new populations have all been shown to be strongly influenced by the match between the host plant composition of otherwise suitable habitat patches and the host plant preference of migrating butterflies. The evolutionary consequences of such biased migration and colonization with respect to butterfly phenotypes might differ depending on spatial configuration and plant species composition of the patches in heterogeneous patch networks. Using a spatially realistic individual-based model we show that the model-predicted evolution of host plant preference due to biased migration explains a significant amount of the observed variation in host plant use among metapopulations living in dissimilar networks. This example illustrates how the ecological extinction-colonization dynamics may be linked with the evolutionary dynamics of life history traits in metapopulations.     

Testing for mating isolation between ecotypes: laboratory experiments with lake, stream and hybrid stickleback

Mating isolation is a frequent contributor to ecological speciation – but how consistently does it evolve as a result of divergent selection? We tested for genetically based mating isolation between lake and stream threespine stickleback (Gasterosteus aculeatus L.) from the Misty watershed, Vancouver Island, British Columbia. We combined several design elements that are uncommon in the studies of stickleback mate choice: (i) we used second‐generation laboratory‐reared fish (to reduce environmental and maternal effects), (ii) we allowed for male–male competitive interactions (instead of the typical no‐choice trials) and (iii) we included hybrids along with pure types. Males of different types (Lake, Inlet, hybrid) were paired in aquaria, allowed to build nests and then exposed sequentially to females of all three types. We found that Lake and Inlet males differed in behaviours thought to influence stickleback mate choice (inter‐ and intra‐sexual aggression, display and nest activities), whereas hybrids were either intermediate or apparently ‘inferior’ in these behaviours. Despite these differences, Lake and Inlet fish did not mate assortatively and hybrid males did not have a mating disadvantage. Our study reinforces the noninevitability of mating isolation evolving in response to ecological differences and highlights the need to further investigate the factors promoting and constraining progress towards ecological speciation.