GROWTH AND MATING OF GONZUM PECTORALE (VOLVOCALES) IN DEFINED MEDIA1

Growth in defined media of 32 populations of Gonium pectorale was studied to learn more about the sexual isolation reported for this species. The 23 populations containing both mating types (+ & -) were also studied for the ability to form zygotes in defined media. A preliminary study showed that some populations grew and reproduced sexually in a defined mineral medium, whereas others appeared to require exogenous organic materials for growth and/or zygote formation. The diverse reactions exhibited by the populations indicate physiological races which may explain, in part, the occurrence of sexual isolation.     

Pre-mating isolation is determined by larval rearing substrates in cactophilicDrosophila mojavensis. II. Effects of larval substrates on time to copulation,mate choice and mating propensity

Summary It has been hypothesized that reproductive character displacement has evolved in mainland Sonora, Mexico populations of cactophilicD. mojavensis due to the presence of a sympatric sibling speciesD. arizonae. In laboratory tests using ancestral Baja California populations and derived, sympatric mainland populations, asymmetrical sexual isolation has been observed among populations ofD. mojavensis where mainland females discriminate against Baja males. Effects of different pre-adult rearing environments on adult mating behaviour were assessed by comparing fermenting cactus tissues like those used in nature for breeding with laboratory media because previous studies have employed synthetic growth media for fly growth and development. Significant behavioural isolation was evident in all cases when larvae were reared on laboratory food, but was non-significant when flies were reared on fermenting cactus, except for the cactus used by most mainland populations, consistent with previous studies. Time to copulation of Baja females was greater than mainland females over all substrates, but male time to copulation did not differ between populations. Time to copulation for both sexes was significantly greater when flies were reared on laboratory food with one exception. The degree of behavioural isolation was weakly correlated with time to copulation across food types (Spearman rank correlation = 0.58,p = 0.099). Therefore, use of laboratory media in this and previous studies exaggerated adult pre-mating isolation and time to copulation in comparison to natural breeding substrates. These experiments suggest that a change in host substrates by saprophagous insects (where chemical differences exist between hosts) may have subtle effects on mating behaviour in a manner which promotes low levels of sexual isolation as a by-product of their utilization of a particular substrate during larval development. ForD. mojavensis, these results suggest that over evolutionary time, radiation into a new environment (from Baja to the mainland) allowed utilization of new host plants that may have incidentally promoted the sexual isolation patterns that have been observed within this species.See Etges (1992) for the first paper in this series.     

The effect of metals and alcohol on sexual isolation in Drosophila melanogaster

Strong sexual isolation established between D. melanogaster long-term cage populations (originated from common parents and being under selection pressure since 1972) is maintained (with a tendency to increase) for twelve years after the origin of the populations. The sexual isolation is also maintained when the populations are kept in common conditions for about two years, while it dramatically decreases when the populations live on a food medium supplemented with strong chemical selective factors, such as various metals or ethanol. Seasonal or geographical studies of sexual isolation between natural and our cage populations did not reveal significant deviation from random matings. The genetic nature of sexual isolation is discussed.     

Sexual Isolation Between Drosophila Melanogaster, D. Simulans and D. Mauritiana: Sex and Species Specific Discrimination

The sexual isolation among the related species Drosophila melanogaster, D. simulans and D. mauritiana is asymmetrical. While D. mauritiana males mate well with both D. melanogaster and D. simulans females, females of D. mauritiana discriminate strongly against males of these two species. Similarly, D. simulans males mate with D. melanogaster females but the reciprocal cross is difficult. Interspecific crosses between several populations of the three species were performed to determine if (i) males and females of the same species share a common sexual isolation genetic system, and (ii) males (or females) use the same genetic system to discriminate against females (or males) of the other two species. Results indicate that although differences in male and female isolation depend on the populations tested, the isolation behaviour between a pair of species is highly correlated despite the variations. However, the rank order of the isolation level along the populations was not correlated in both sexes, which suggests that different genes act in male and female sexual isolation. Neither for males nor for females, the isolation behaviour of one species was paralleled in the other two species, which indicates that the genetic systems involved in this trait are species-pair specific. The implications of these results are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

SEXUAL POPULATIONS OF GONIUM PECTORALE (VOLVOCALES)

An investigation of 33 sexual populations of Gonium pectorale has shown sexual isolation not to be as prevalent as previously reported. Only one population was homothallic. Populations from different geographic areas are, in general, sexually compatible. Three groups are discernible based on the degree of intercrossing—the first containing the majority of populations, the second containing only 5 populations, and the third consisting of populations intercrossing with no others. The factors possibly involved in the variable fertility of the populations are discussed. These include the geological history of the collecting areas, migration routes of waterfowl, transport of viable cells by passive means, and cultural conditions (such as pH, temperature, media) used in this investigation.     

Patterns of reproductive isolation within and between two Lygaeus species characterized by sexual conflicts over mating

Theory suggests that, under some circumstances, sexual conflict over mating can lead to divergent sexually antagonistic coevolution among populations for traits associated with mating, and that this can promote reproductive isolation and hence speciation. However, sexual conflict over mating may also select for traits (e.g. male willingness to mate) that enhance gene flow between populations, limiting population divergence. In the present study, we compare pre‐ and post‐mating isolation within and between two species characterized by male–female conflict over mating rate. We quantify sexual isolation among five populations of the seed bug Lygaeus equestris collected from Italy and Sweden, and two replicates of a population of the sister‐species Lygaeus simulans, also collected from Italy. We find no evidence of reproductive isolation amongst populations of L. equestris, suggesting that sexual conflict over mating has not led to population divergence in relevant mating traits in L. equestris. However, there was strong asymmetric pre‐mating isolation between L. equestris and L. simulans: male L. simulans were able to mate successfully with female L. equestris, whereas male L. equestris were largely unable to mate with female L. simulans. We found little evidence for strong post‐mating isolation between the two species, however, with hybrid F₂ offspring being produced. Our results suggest that sexual conflict over mating has not led to population divergence, and indeed perhaps supports the contrary theoretical prediction that male willingness to mate may retard speciation by promoting gene flow.     

A STOCHASTIC SIMULATION STUDY ON SPECIATION BY SEXUAL SELECTION

A two-locus multi-allele sexual isolation model incorporating mutation and genetic drift which was first proposed by Nei et al. (1983) is studied here. One locus controls the male mating character, and the other controls female receptivity. All females are assumed to have equal mating success. Therefore, the frequencies of female receptivity alleles are changed by mutation, drift, and hitchhiking with male character alleles. Without hitchhiking, development of sexual isolation between allopatric populations proceeds faster in smaller populations, as expected. The hitchhiking effect, by triggering the mutual reinforcement of mating behavior of both sexes (or the runaway process, Fisher [1958]), speeds up the evolution of sexual isolation significantly. For populations with 2Nv ≤ 0.2 (N = population size, ν = mutation rate), the rates of divergence all approach the maximum possible rate. Sympatric sexual isolation develops quite frequently if two favorable conditions are met: 1) There is no selection on female phenotype (except in some limited cases), and 2) The population size is large enough to carry several female receptivity alleles. Because of stochastic factors, these alleles may lead to the formation of two discrete groups of females, each group receptive to males of different mating characters. The formation of sympatric sexually-isolated groups is also aided significantly, at the incipient stage, by the runaway process.     

Sexual Activity and Reproductive Isolation Between Age‐specific Selected Populations of Seed Beetle

We evaluated the degree of reproductive isolation between laboratory populations of the seed beetle (Acanthoscelides obtectus) selected to reproduce early (E) or late (L) in life, where different levels of sexual activity and sexual discrimination have been detected. We found a significant level of behavioral isolation among populations within the E selection regime in which beetles showed enhanced early‐life fitness traits and low sexual activity. In contrast, substantially higher levels of sexual activity and an indiscriminate mating system inhibited rather than promoted pre‐zygotic isolation between the L populations. Our results indicate that the study of sexual activity levels may be crucial for understanding the first steps in the pre‐zygotic isolation among allopatric populations subjected to uniform selection.     

Reproductive character displacement of female mate preferences for male cuticular hydrocarbons in Drosophila subquinaria

Several lines of evidence implicate sexual isolation in both initiating and completing the speciation process. Although its existence is straightforward to demonstrate, understanding the evolution of sexual isolation requires identifying the underlying phenotypes responsible so that we can determine how these have diverged. Here, we study geographic variation in female mate preferences for male sexual displays in the fly Drosophila subquinaria. Female D. subquinaria that are sympatric with its sister species D. recens discriminate strongly against both D. recens and allopatric conspecific males, whereas females from allopatric populations do not. Furthermore, female mate preferences target at least in part a suite of cuticular hydrocarbons (CHCs) in males and geographic variation in CHCs mirrors the pattern of mate discrimination. In this study, we quantify female mate preferences for male CHCs from populations that span the geographic range of D. subquinaria. We find that the direction of linear sexual selection varies significantly between populations that are sympatric versus allopatric with D. recens in a pattern of reproductive character displacement. Differences in preference partially align with existing differences in CHCs and patterns of sexual isolation, although discrepancies remain that suggest the involvement of additional traits and/or more complex, nonlinear preference functions.     

Natural selection and divergence in mate preference during speciation

Sexual isolation can evolve due to natural selection against hybrids (reinforcement). However, many different forms of hybrid dysfunction, and selective processes that do not involve hybrids, can contribute to the evolution of sexual isolation. Here we review how different selective processes affect the evolution of sexual isolation, describe approaches for distinguishing among them, and assess how they contribute to variation in sexual isolation among populations of Timema cristinae stick-insects. Pairs of allopatric populations of T. cristinae living on different host-plant species exhibit greater sexual isolation than those on the same host, indicating that some sexual isolation has evolved due to host adaptation. Sexual isolation is strongest in regions where populations on different hosts are in geographic contact, a pattern of reproductive character displacement that is indicative of reinforcement. Ecological costs to hybridization do occur but traits under ecological selection (predation) do not co-vary strongly with the probability of between-population mating such that selection on ecological traits is not predicted to produce a strong correlated evolutionary response in mate preference. Moreover, F1 hybrid egg inviability is lacking and the factors contributing to reproductive character displacement require further study. Finally, we show that sexual isolation involves, at least in part, olfactory communication. Our results illustrate how understanding of the evolution of sexual isolation can be enhanced by isolating the roles of diverse ecological and evolutionary processes.     

DISENTANGLING THE EFFECTS OF MATING PROPENSITY AND MATING CHOICE IN DROSOPHILA

Incipient sexual isolation between genotypes, lines, or populations of the same species is commonly measured in Drosophila by choice tests. Results of these tests are known to be influenced, in an undetermined manner, by the mating propensity of competitors and by discriminatory factors during courtship. We have approached the problem by measuring male and female propensities in separate, independent tests, and by examining whether these estimates could explain the results of the choice tests. First, male and female choice tests were used to measure sexual isolation between populations of Drosophila melanogaster and between populations of D. simulans. Significant deviations from random mating occurred in 31 out of 48 tests, in agreement with the propensity values of the tested genotypes. We conclude that mating propensity instead of discrimination is directly involved in the estimation of sexual isolation in our populations, and advise against the application of male and female choice tests to assess intraspecific isolation without a proper knowledge of the mating propensities of competing individuals. Second, multiple choice tests were used to assess isolation between D. melanogaster populations. In examining the dynamics of matings throughout the test, we show that if competing individuals differ in mating propensities and tests are long enough to allow most matings to happen, a spurious sexual isolation can appear. We recommend that multiple choice tests be terminated once 50 percent of matings had been observed.     

Sexual isolation with and without ecological isolation in marine isopods Jaera albifrons and J. praehirsuta

Sexual barriers associated with mate choice are often found to be associated with some level of ecological isolation between species. The independence and relative strength of sexual isolation are thus difficult to assess. Here, we take advantage of a pair of marine isopod species (Jaera albifrons and J. praehirsuta) that show sexual isolation and coexist in populations where they share the same microhabitat or not (i.e. without or with ecological isolation). We estimated the strength of sexual isolation between J. albifrons and J. praehirsuta using no‐choice trials and a multiple‐choice experimental population. We found that sexual isolation is strong in both the presence and the absence of ecological isolation, but that it is asymmetric and fails to prevent interspecific gene flow entirely. First‐generation intrinsic post‐zygotic barriers were low, and there was no sexual isolation within J. praehirsuta across habitats. The J. albifrons/J. praehirsuta species pair thus provides an example where the role of sexual isolation as a barrier to gene flow (a) does not depend upon current ecological isolation, (b) seems to have evolved independently of local ecological conditions, but (c) is insufficient to complete speciation entirely on its own.     

Reproductive strategies and isolation‐by‐demography in a marine clonal plant along an eutrophication gradient

Genetic diversity in clonal organisms includes two distinct components, (i) the diversity of genotypes or clones (i.e. genotypic richness) in a population and (ii) that of the alleles (i.e. allelic and gene diversity within populations, and differentiation between populations). We investigated how population differentiation and genotypic components are associated across a gradient of eutrophication in a clonal marine plant. To that end, we combined direct measurements of sexual allocation (i.e. flower and seed counts) and genotypic analyses, which are used as an estimator of effective sexual reproduction across multiple generations. Genetic differentiation across sites was also modelled according to a hypothesis here defined as isolation‐by‐demography, in which we use population‐specific factors, genotypic richness and eutrophication that are hypothesized to affect the source‐sink dynamics and thus influence the genetic differentiation between a pair of populations. Eutrophic populations exhibited lower genotypic richness, in agreement with lower direct measurements of sexual allocation and contemporaneous gene flow. Genetic differentiation, while not explained by distance, was best predicted by genotypic richness and habitat quality. A multiple regression model using these two predictors was considered the best model (R² = 0.43). In this study, the relationship between environment and effective sexual–asexual balance is not simply (linearly) predicted by direct measurements of sexual allocation. Our results indicate that population‐specific factors and the isolation‐by‐demography model should be used more often to understand genetic differentiation.     

The role of sexual isolation during rapid ecological divergence: Evidence for a new dimension of isolation in Rhagoletis pomonella

The pace of divergence and likelihood of speciation often depends on how and when different types of reproductive barriers evolve. Questions remain about how reproductive isolation evolves after initial divergence. We tested for the presence of sexual isolation (reduced mating between populations due to divergent mating preferences and traits) in Rhagoletis pomonella flies, a model system for incipient ecological speciation. We measured the strength of sexual isolation between two very recently diverged (~170 generations) sympatric populations, adapted to different host fruits (hawthorn and apple). We found that flies from both populations were more likely to mate within than between populations. Thus, sexual isolation may play an important role in reducing gene flow allowed by early-acting ecological barriers. We also tested how warmer temperatures predicted under climate change could alter sexual isolation and found that sexual isolation was markedly asymmetric under warmer temperatures – apple males and hawthorn females mated randomly while apple females and hawthorn males mated more within populations than between. Our findings provide a window into the early speciation process and the role of sexual isolation after initial ecological divergence, in addition to examining how environmental conditions could shape the likelihood of further divergence.     

Strength of sexual and postmating prezygotic barriers varies between sympatric populations with different histories and species abundances

The impact of different reproductive barriers on species or population isolation may vary in different stages of speciation depending on evolutionary forces acting within species and through species’ interactions. Genetic incompatibilities between interacting species are expected to reinforce prezygotic barriers in sympatric populations and lead to cascade reinforcement between conspecific populations living within and outside the areas of sympatry. We tested these predictions and studied whether and how the strength and target of reinforcement between Drosophila montana and Drosophila flavomontana vary between sympatric populations with different histories and species abundances. All barriers between D. montana females and D. flavomontana males were nearly complete, while in the reciprocal cross strong postzygotic isolation was accompanied by prezygotic barriers whose strength varied according to population composition. Sexual isolation between D. flavomontana females and D. montana males was increased in long‐established sympatric populations, where D. flavomontana is abundant, while postmating prezygotic (PMPZ) barriers were stronger in populations where this species is a new invader and still rare and where female discrimination against heterospecific males was lower. Strengthening of sexual and PMPZ barriers in this cross also induced cascade reinforcement of respective barriers between D. flavomontana populations, which is a classic signature of reinforcement process.     

Reinforcement shapes clines in female mate discrimination in Drosophila subquinaria

Reinforcement of species boundaries may alter mate recognition in a way that also affects patterns of mate preference among conspecific populations. In the fly Drosophila subquinaria, females sympatric with the closely related species D. recens reject mating with heterospecific males as well as with conspecific males from allopatric populations. Here, we assess geographic variation in behavioral isolation within and among populations of D. subquinaria and use cline theory to understand patterns of selection on reinforced discrimination and its consequences for sexual isolation within species. We find that selection has fixed rejection of D. recens males in sympatry, while significant genetic variation in this behavior occurs within allopatric populations. In conspecific matings sexual isolation is also asymmetric and stronger in populations that are sympatric with D. recens. The clines in behavioral discrimination within and between species are similar in shape and are maintained by strong selection in the face of gene flow, and we show that some of their genetic basis may be either shared or linked. Thus, while reinforcement can drive extremely strong phenotypic divergence, the long‐term consequences for incipient speciation depend on gene flow, genetic linkage of discrimination traits, and the cost of these behaviors in allopatry.     

Geographical variation of chromosomal structure in Drosophila gasici

Summary Drosophila gasici Brncic 1957, is a neotropical species found in several parts of the Andes Mountain System. By means of the analysis of their external characteristics, chromosomes and hybridization test they have been included in the mesophragmatica group of species of the sub-genus Drosophila.The present paper describes the mitotic and polytene chromosomes of D. gasici from samples of natural populations collected at Bogotá (Colombia), Cochabamba (Bolivia), Arica (Chile) and San Luis (Argentina). The comparative study of all these populations has disclosed that the species has split in well defined geographic races. The Colombian and Chilean flies differ from those living in Bolivia and Argentina by three independent inversions in chromosome I (the sexual pair). The only polymorphic populations seem to be the Chilean ones which exhibit two inversions in the second chromosome, besides the Standard gene arrangement. All the other populations are homozygous for all their chromosomal sequences. Studies on reproductive isolation have demonstrated that there is some sexual discrimination between the Colombian and Chilean flies in respect to the Bolivian and Argentinean ones.The populational structure of D. gasici is in contrast to that observed in the other six species belonging to the mesophragmatica group in which there are no good evidences of geographical variations at the chromosomal level.This article is dedicated to Professor Hans Bauer on the occasion of his 60th birthday.     

Revealing the mechanisms of sexual isolation in a case of sympatric and parallel ecological divergence

Two ecotypes of a marine intertidal snail (Littorina saxatilis), living at different microhabitats and shore levels, have evolved in sympatry and in parallel across the Galician rocky shore. These ecotypes differ in many traits (including size) due to differential adaptation. They meet, mate assortatively, and partially hybridize at the mid shore where the two microhabitats overlap. The partial sexual isolation observed is claimed to be a side‐effect of the size differences between ecotypes combined with a size assortative mating found in most populations of this species. We investigated this hypothesis using three complementary experimental approaches. First, we investigated which of the different shell variables contributed most to the variation in individual sexual isolation in the field by using two new statistics developed for that purpose: (1) pair sexual isolation and (2) r_i, which is based on the Pearson correlation coefficient. We found that size is the most important trait explaining the sexual isolation and, in particular, the males appear to be the key sex contributing to sexual isolation. Second, we compared the size assortative mating between regions: exposed rocky shore populations from north‐westwern Spain (showing incomplete reproductive isolation due to size assortative mating) and protected Spanish and Swedish populations (showing size assortative mating but not reproductive isolation between ecomorphs). Most of the variation in size assortative mating between localities was significantly explained by the within‐population level of variation on size. Third, we performed a laboratory male choice experiment, which further suggested that the choice is made predominantly on the basis of size. These results confirm the mechanism proposed to explain the sexual isolation in the Galician hybrid zone and thus support this case as a putative example of parallel incipient speciation. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 94 , 513–526.     

THE EVOLUTION OF ASYMMETRY IN SEXUAL ISOLATION: A MODEL AND A TEST CASE

We constructed a model for the evolution of sexual isolation by extending Lande's (1981) model of sexual selection. The model predicts that asymmetric sexual isolation is a transient phenomenon, characteristic of intermediate stages of divergence in sexually selected traits. Unlike the Kaneshiro (1976, 1980) proposal, our model does not depend upon drift and the loss of courtship elements to produce asymmetries in sexual isolation. According to our model, the direction of evolution cannot be predicted from asymmetry in sexual isolation. We tested some features of the model using data from an experimental study of sexual isolation in the salamander Desmognathus ochrophaeus. We tested for sexual isolation between 12 allopatric populations and found significant asymmetry in sexual isolation in about a quarter of the test cases. The highest degrees of asymmetry were associated with intermediate levels of divergence. A curvilinear relationship between isolation asymmetry and divergence was predicted by our model and was supported by statistical analysis of the salamander data.     

Effect of chromosome arrangements on mate recognition system leading to behavioral isolation in <Emphasis Type="Italic">Drosophila ananassae</Emphasis>

The mechanisms of speciation that appear in the early stages of reproductive isolation has been of recent interest to evolutionary biologists. Experiments were conducted to study behavioral isolation between karyotypically different homozygous strains derived from natural populations of Drosophila ananassae. Three mass cultures stocks established from flies collected from natural populations were employed and homozygous stocks (ST/ST and AL/AL) were made through selection for homozygosity. By employing male-choice technique, mating success was scored by direct observation in the Elens–Wattiaux mating chamber. There is preference for homogamic matings in all the three populations and the differences between homogamic and heterogamic matings are statistically significant in two populations (Lucknow and Varanasi). These findings provide evidence that there is incipient sexual isolation between karyotypically different strains of D. ananassae derived from natural populations which shows that chromosome arrangements may affect the mate recognition system in D. ananassae.