Drosophila melanogaster males respond differently at the behavioural and genome‐wide levels to Drosophila melanogaster and Drosophila simulans females

Drosophila melanogaster are found in sympatry with Drosophila simulans, and matings between the species produce nonfertile hybrid offspring at low frequency. Evolutionary theory predicts that females choose mates, so males should alter their behaviour in response to female cues. We show that D. melanogaster males quickly decrease courtship towards D. simulans females. Courtship levels are reduced within 5 min of exposure to a heterospecific female, and overall courtship is significantly lower than courtship towards conspecific females. To understand changes at the molecular level during mate choice, we performed microarray analysis on D. melanogaster males that courted heterospecific D. simulans females and found nine genes have altered expression compared with controls. In contrast, males that court conspecific females alter expression of at least 35 loci. The changes elicited by conspecific courtship likely modulate nervous system function to reinforce positive conspecific signals and dampen the response to heterospecific signals.     

The magnitude of behavioral isolation is affected by characteristics of the mating community

Gene exchange between species occurs in areas of secondary contact, where two species have the opportunity to hybridize. If heterospecific males are more common than conspecific males, females will experience more encounters with males of other species. These encounters might increase the likelihood of heterospecific matings, and lead to the production of hybrid progeny. I studied the mating behavior of two pairs of sibling species endemic to Africa: Drosophila yakuba/Drosophila santomea and Drosophila simulans/Drosophila sechellia. Drosophila yakuba and D. simulans are cosmopolitan species widely distributed in the African continent, while D. santomea and D. sechellia are island endemics. These pairs of species hybridize in nature and have the potential to exchange genes in natural conditions. I used these two pairs of Drosophila species, and constructed mating communities of different size and different heterospecific:conspecific composition. I found that both the total number of potential mates and the relative frequency of conspecific versus heterospecific males affect female mating decisions in the cosmopolitan species but not in the island endemics. These results suggest that the population characteristics, in which mating occurs, may affect the magnitude of premating isolation. Community composition might thus facilitate, or impair, gene flow between species.     

Absence of social facilitation of courtship in the wolf spider, Schizocosa ocreata (Hentz) (Araneae: Lycosidae)

Males of many animal species are reproductively limited by the difficulty and time costs of finding mates. Males of such species should be selected to take advantage of any cues that might reveal the location of prospective mates. Cues to female location are not restricted to those produced by females, but might also include the highly apparent courtship displays of males that have already found a female. By “eavesdropping” on these courting rivals, initiating sexual displays when courting rivals are detected (i.e., social facilitation of displays); males might effectively exploit the mate-searching efforts of their rivals. We tested the possibility that male Schizocosa ocreata wolf spiders exhibit social facilitation of courtship behaviors using a combination of live behavioral trials and video playback with single stimulus presentations. When exposed to visual cues from another male, male S. ocreata can discern the presence of another individual whether that individual is courting or not. However, we found no evidence of social facilitation of courtship or chemoexploratory behaviors in response to seismic or visual cues presented in isolation or combined. While complex, multimodal, male courtship signals are important in mate choice by female S. ocreata, males do not appear to use these cues to socially facilitate their own courtship.     

SIMULATING RANGE EXPANSION: MALE SPECIES RECOGNITION AND LOSS OF PREMATING ISOLATION IN DAMSELFLIES

Prolonged periods of allopatry might result in loss of the ability to discriminate against other formerly sympatric species, and can lead to heterospecific matings and hybridization upon secondary contact. Loss of premating isolation during prolonged allopatry can operate in the opposite direction of reinforcement, but has until now been little explored. We investigated how premating isolation between two closely related damselfly species, Calopteryx splendens and C. virgo , might be affected by the expected future northward range expansion of C. splendens into the allopatric zone of C. virgo in northern Scandinavia. We simulated the expected secondary contact by presenting C. splendens females to C. virgo males in the northern allopatric populations in Finland. Premating isolation toward C. splendens in northern allopatric populations was compared to sympatric populations in southern Finland and southern Sweden. Male courtship responses of C. virgo toward conspecific females showed limited geographic variation, however, courtship attempts toward heterospecific C. splendens females increased significantly from sympatry to allopatry. Our results suggest that allopatric C. virgo males have partly lost their ability to discriminate against heterospecific females. Reduced premating isolation in allopatry might lead to increased heterospecific matings between taxa that are currently expanding and shifting their ranges in response to climate change.     

No evidence for learned mating discrimination in male Drosophila pseudoobscura

Background  

Since females often pay a higher cost for heterospecific matings, mate discrimination and species recognition are driven primarily by female choice. In contrast, frequent indiscriminate matings are hypothesized to maximize male fitness. However, recent studies show that previously indiscriminate males (e.g., Drosophila melanogaster and Poecilia reticulata) can learn to avoid heterospecific courtship. This ability of males to discriminate against heterospecific courtship may be advantageous in populations where two species co-occur if courtship or mating is costly.     

Alternative reproductive tactics in male freshwater fish influence the accuracy of species recognition

Sexual conflict can result in coercive mating. Because males bear low costs of heterospecific mating, coercive males may engage in misdirected mating attempts toward heterospecific females. In contrast, sexual selection through consensual mate choice can cause mate recognition cues among species to diverge, leading to more accurate species recognition. Some species show both coercive mating and mate choice‐associated courtship behaviors as male alternative reproductive tactics. We hypothesized that if the selection pressures on each tactic differ, then the accuracy of species recognition would also change depending on the mating tactic adopted. We tested this hypothesis in the guppy (Poecilia reticulata) and mosquitofish (Gambusia affinis) by a series of choice experiments. Poecilia reticulata and G. affinis males both showed imperfect species recognition and directed all components of mating behavior toward heterospecific females. They tended to direct courtship displays more frequently toward conspecific than heterospecific females. With male P. reticulata, however, accurate species recognition disappeared when they attempted coercive copulation: they directed coercions more frequently toward heterospecific females. We also found that heterospecific sexual interaction had little effect on the fecundity of gravid females, which suggests that prepregnancy interactions likely underpin the exclusion of G. affinis by P. reticulata in our region.     

ACOUSTIC COMMUNICATION AND MATING BEHAVIOUR IN WATER BUGS OF THE GENUS MICRONECTA

ABSTRACT

The role of male sounds in attracting females and in mating was investigated in the three most common species of Micronecta found in ponds 60 km NE of Melbourne, Australia: M. concordia, M. tasmanica and M. robusta. In playback experiments using recorded male signals, females were attracted to signals of conspecific males, in preference to signals of heterospecific sympatric males. Studies of mating behaviour, using video recording, showed that signals were obligatory for mating. These findings strongly suggest that acoustic signalling is important in reproductive isolation in Micronecta. Comparisons between matings and rejected matings showed that, within each species, copulation only occurred following a certain minimum number of acoustic signals with more pulse-trains than the mean for the species. Low amplitude sounds (after signals) were also important in courtship, immediately preceding mating. No sounds occurred during copulation.     

Possible ecological consequences of heterospecific mating behavior in two tetranychid mites

Interspecific mating between the two-spotted spider mite,Tetranychus urticae Koch, and the Banks grass mite,Oligonychus pratensis (Banks), was documented using laboratory populations. The incidence of mating betweenT. urticae males andO. pratensis females was 26.0%, while that for the reciprocal mating was 18.8%. The incidence of mating was affected by both male and female species. Such matings may have several important ecological consequences. Interspecific matings resulted in all-male progenies. Thus, progeny sex ratios may be distorted by misdirected mating behavior. In addition, heterospecific mating resulted in lower fecundity than conspecific matings in the two-spotted spider mite, although not in the Banks grass mite. Aerial dispersal behavior of the two-spotted spider mite was also affected. Under crowded conditions and deteriorating resource quality, female mites exhibit an aerial dispersal posture that helps them to become airborne, and allows them to disperse long distances. Forty-two percent ofT. urticae females that mated with conspecific males exhibited this dispersal behavior, compared to only 3.6% for virgin females. The incidence of aerial dispersal behavior for females that mated with heterospecific males was intermediate (27.3%). The effects of these behavioral alterations on male and female fitness may depend on the population structure and resource distribution.     

Body Size and the Maintenance of Reproductive Isolation in Stickleback,Genus Gasterosteus

The blackspotted stickleback Gasterosteus wheatlandi and the widely studied threespine stickleback G. aculeatus are sympatric throughout the former’s range and share many aspects of life history and reproductive behaviour. These two species differ significantly in size, with G. wheatlandi of both sexes measured at approximately 60% of the standard length of their G. aculeatus counterparts. This study concentrated on G. wheatlandi courtship behaviour and investigated its role in the maintenance of reproductive isolation with G. aculeatus. Specifically, the roles that (1) female body size plays in influencing male courtship preferences and (2) male body size and behaviour play in female courtship preferences were investigated through dummy and live conspecific and heterospecific stimulus presentations. Male G. wheatlandi courtship preferences are consistent with previously described patterns for G. aculeatus. Males of both species preferentially approach and court the larger of two simultaneously presented live or dummy females. Thus, the smaller G. wheatlandi males are indiscriminate with respect to assortative mate choice; not only preferring to approach and court more fecund conspecific females but, more significantly, G. aculeatus‐sized females. In contrast, females of both species demonstrate strong assortative courtship preferences. When presented with pairs of flask‐enclosed males, females of both species preferentially orient and court the conspecific male over the heterospecific. Similarly, when presented with a conspecific male and a heterospecific male presented singly, females prefer to enter the nest of the conspecific. Systematic analysis of the interactions between these pairs of fish (one male, one female) demonstrates that the breakdown of courtship in heterospecific courtship occurs late in the courtship sequence when the widely differing forms of male leading behaviour results in drastically differing female responses. I suggest that, as previously described in G. aculeatus, the supernormality effect plays a significant role in mediating adaptive mate choice behaviour in G. wheatlandi. However, the added element of a larger sympatric species introduces a possible cost in time and energy devoted to courting heterospecific, and sympatric, females that the larger G. aculeatus do not likely incur. There is substantial evidence from many sympatric G. aculeatus species pairs that there is assortative mate choice based on size and/or courtship behaviour. Courtship trials suggest a more pervasive role for females in assortative mate choice. Whether it is male body size per se, or in combination with behaviour, morphology or other cues, is unresolved in the present study.     

Reproductive interference in two ground-hopper species: testing hypotheses of coexistence in the field

Similar to resource competition, reproductive interference may hamper the coexistence of closely related species. Species that utilize similar signal channels during mate finding may face substantial fitness costs when they come into contact and demographic displacement of the inferior species (sexual exclusion) is a likely outcome of such interactions. The two ground‐hopper species Tetrix ceperoi and Tetrix subulata broadly overlap in their ranges and general habitat requirements, but rarely co‐occur on a local scale. Results from laboratory and field experiments suggest that this mosaic pattern of sympatry might be influenced by reproductive interference. Here, we examine the significance of sexual interactions for these species in the field and test hypotheses on mechanisms of coexistence. Our results show that heterospecific sexual interactions also occur under field conditions, but in contrast to the experiments T. ceperoi was not the inferior species. The number of male mating attempts of both species was strongly correlated with encounter frequencies. Males discriminated between the sexes but not between the species, suggesting an incomplete mate recognition system in both species. The analysis of microhabitat preferences and spatial distribution revealed that habitat partitioning is not a suitable mechanism of coexistence in this system. Instead, the costs of reproductive interference are substantially mitigated by different niche breadths leading to different degrees of aggregation. Despite a considerable niche overlap T. ceperoi displayed a stronger preference for bare ground and occurred more aggregated than T. subulata, which had a broader niche. These differences may reduce the frequencies of heterospecific encounters and interactions in the field. Our results demonstrate that coexistence in the presence of reproductive interference is comparable to resource competition, being strongly influenced by ecological traits of the involved species, such as niche breadth and dispersion pattern.     

Selective phonotaxis in Tettigonia cantans and T. viridissima in song recognition and discrimination

The selectivity of female phonotaxis in Tettigonia cantans and T. viridissima was investigated on a Kramer treadmill, with respect to the specific differences in temporal pattern and spectrum of the songs of both species. In choice situations, both species preferred the conspecific song over the heterospecific one. The courses of both species were deflected by about 15–20° from the position of the conspecific song, that of T. viridissima being away from, that of T. cantans in the direction of the heterospecific song. In no-choice situations, song models with the temporal pattern of T. cantans did not attract T. viridissima. Models with the conspecific time pattern but heterospecific spectrum were as attractive as the conspecific model. In contrast, T. cantans was attracted by T. viridissima song presented alone. In choice situations, either spectral or temporal differences were sufficient for discrimination. The preference for the conspecific model gradually disappeared with stepwise reduction of its intensity and was reversed at −12 dB. Acoustic communication alone can serve species isolation in T. viridissima; however, premating isolation in T. cantans must involve other mechanisms. The orientation during the choice situations suggests a serial processing of song recognition and localization for the Tettigonia species. Accepted: 13 December 1997     

The effects of heterospecific mating frequency on the strength of cryptic reproductive barriers

Heterospecific mating frequency is critical to hybrid zone dynamics and can directly impact the strength of reproductive barriers and patterns of introgression. The effectiveness of post‐mating prezygotic (PMPZ) reproductive barriers, which include reduced fecundity via heterospecific matings and conspecific sperm precedence, may depend on the number, identity and order of mates. Studies of PMPZ barriers suggest that they may be important in many systems, but whether these barriers are effective at realistic heterospecific mating frequencies has not been tested. Here, we evaluate the strength of cryptic reproductive isolation in two leaf beetles (Chrysochus auratus and C. cobaltinus) in the context of a range of heterospecific mating frequencies observed in natural populations. We found both species benefited from multiple matings, but the benefits were greater in C. cobaltinus and extended to heterospecific matings. We found that PMPZ barriers greatly limited hybrid production by C. auratus females with moderate heterospecific mating frequencies, but that their effectiveness diminished at higher heterospecific mating frequencies. In contrast, there was no evidence for PMPZ barriers in C. cobaltinus females at any heterospecific mating frequency. We show that integrating realistic estimates of cryptic isolation with information on relative abundance and heterospecific mating frequency in the field substantially improves our understanding of the strong directional bias in F1 production previously documented in the Chrysochus hybrid zone. Our results demonstrate that heterospecific mating frequency is critical to understanding the impact of cryptic post‐copulatory barriers on hybrid zone structure and dynamics, and that future studies of such barriers should incorporate field‐relevant heterospecific mating frequencies.     

Reproductive interference by male Drosophila subobscura on female D. persimilis: A laboratory experiment

While females often reject courtship attempts by heterospecific males, reproductive interference by harassment from such males can nonetheless occur, potentially reducing female fitness. Such effects may be profound following a range expansion, when males from a new species may suddenly encounter (and perhaps even become abundant relative to) females of related native species. Drosophila subobscura recently invaded North America and may impact native species through reproductive interference and other processes. We test for the potential for reproductive interference by D. subobscura males on D. persimilis females in the laboratory. D. subobscura males aggressively copulated with D. persimilis females, including many females that exhibit rejection behaviors. Despite females attempting to dismount the males, the heterospecific copulations are on average longer than conspecific copulations, and females exhibit some reluctance to remate with conspecific males following this harassment. Females confined with both conspecific and heterospecific males produce fewer adult progeny than those with either conspecific males only or with conspecific males and distantly related D. simulans males that do not engage in female harassment. Overall, our results illustrate how reproductive interference by an invasive species can have negative effects on resident natural populations.     

SEX DIFFERENCES IN MATE RECOGNITION AND CONSPECIFIC PREFERENCE IN SPECIES WITH MUTUAL MATE CHOICE

Sexual isolation is often assumed to arise because choosy females recognize and reject heterospecific males as mates. Yet in taxa in which both males and females are choosy, males might also recognize and reject heterospecific females. Here, we asked about the relative contribution of the sexes to the strong sexual isolation found in limnetic–benthic species pairs of threespine sticklebacks, which show mutual mate choice. We asked whether males and females of the two species recognize conspecifics and also prefer to mate with them. We found evidence for mate recognition by both sexes but only females prefer conspecifics. The nature of male courtship depended on which species of female they were courting, indicating that males recognized conspecific females and differentiated them from heterospecifics. However, males courted both species of females with equal vigor and changed courtship in a manner that would increase the chance of mating with heterospecifics. Females both recognized conspecifics and strongly preferred them. They responded very little to heterospecific male courtship and almost never mated with them. Therefore, males are likely to undermine sexual isolation, but females uphold it. Despite mutual mate choice and mate recognition in both sexes, females are primarily responsible for sexual isolation in these taxa.     

Impact of experimental design on Drosophila sexual isolation studies: direct effects and comparison to field hybridization data

Abstract Many studies of speciation rely critically on estimates of sexual isolation obtained in the laboratory. Here we examine the sensitivity of sexual isolation to alterations in experimental design and mating environment in two sister species of Drosophila, D. santomea and D. yakuba. We use a newly devised measure of mating frequencies that is able to disentangle sexual isolation from species differences in mating propensity. Variation in fly density, presence or absence of a quasi‐natural environment, degree of starvation, and relative frequency of species had little or no effect on sexual isolation, but one factor did have a significant effect: the possibility of choice. Designs that allowed flies to choose between conspecific and heterospecific mates showed significantly more sexual isolation than other designs that did not allow choice. These experiments suggest that sexual isolation between these species (whose ranges overlap on the island of STo Tomé) is due largely to discrimination against D. yakuba males by D. santomea females. This suggestion was confirmed by direct observations of mating behavior. Drosophila santomea males also court D. yakuba females less ardently than conspecific females, whereas neither males nor females of D. yakuba show strong mate discrimination. Thus, sexual isolation appears to be a result of evolutionary changes in the derived island endemic D. santomea. Surprisingly, as reported in a companion paper (Llopart et al. 2005), the genotypes of hybrids found in nature do not accord with expectations from these laboratory studies: all F₁ hybrids in nature come from matings between D. santomea females and D. yakuba males, matings that occur only rarely in the laboratory.     

Effects of Courtship Song in Interspecific Crosses Among the Species of the Drosophila auraria Complex (Diptera: Drosophilidae)

The courtship behavior and the effects of courtship song in inter- and intraspecific crosses were studied in the four sympatric species of the Drosophila auraria complex: D. auraria, D. biauraria, D. subauraria, and D. triauraria. Orientation, tapping, and vibration (the repertoires of male courtship) were observed in both inter- and intraspecific crosses, suggesting that signals from heterospecific females were enough to elicit such male behaviors. The crossability tests with wingless or winged heterospecific males (tests for wing effects) revealed that winged heterospecific males copulated less than wingless ones in all four species but not all the pairwise cases. Since the crossability tests with aristaless females (deaf) or normal females showed essentially the same results as the tests for wing effects, we concluded that the sound produced by wing vibration plays an important role and that the wing movement itself is less important. These findings suggest that courtship songs are of great importance in mate discrimination and the sexual isolation between the species of this complex.     

Agonistic and courtship behaviour patterns in the skink Chalcides viridanus (Fam. Scincidae) from Tenerife

There have been relatively few attempts to quantitatively describe behaviours in scincid lizards. Chalcides viridanus is a small body-sized skink endemic of Tenerife (Canary Islands). We describe and quantify 18 behaviour patterns (both social and agonistic) of this species, some of which have not been described before for other scincids. Video recordings of male–male, female–female, and male–female interactions were made under laboratory conditions, with controlled light–dark cycle and temperature. We describe several agonistic and courtship behaviour patterns. Within the first context, we detected a new agonistic behaviour for a scincid, “Snout to body”, that appeared at the beginning of agonistic sequences; it consisted of each animal placing its snout in contact with the other individual’s lateral side of the body. The amplitude of head movement during “Head bobbing” was lower than that described for many other lizard species. Agonistic behaviours were shown in intrasexual staged encounters both within males and females. The comparison of behaviour patterns of both types of intrasexual encounters showed that females were more active, exhibiting significantly higher frequencies of behaviour than males. Specifically, females showed the “Snout to body” pattern more frequently than males. In male–female encounters we detected courtship and copulation patterns only in April, when males performed “Bites” and “Snout to body” directed at females.     

Behavioral Sequence Leading to Sexual Isolation Between <Emphasis Type="Italic">Drosophila ananassae</Emphasis> and <Emphasis Type="Italic">D. pallidosa</Emphasis>

Drosophila ananassae and D. pallidosa are closely related, sympatric species that lack postmating isolation. Sexual isolation has been considered important in maintaining them as independent species. To clarify the behavioral processes leading to sexual isolation, we analyzed behavioral sequences and examined the effect of courtship song on mating success and on behaviors of both sexes by surgically removing male wings (song generators), female aristae (song receivers), or female wings (means of fluttering). We found that heterospecific courtship songs evoked female wing fluttering, whereas conspecific courtship song did not. Furthermore, female wing fluttering made courting males discontinue courtship. These findings suggest that strong sexual isolation is achieved through the following behavioral sequence: heterospecific song→female wing fluttering→courtship discontinuation.     

Sequential mate choice and sexual isolation in threespine stickleback species

Sequential mate choice strategies predict how females should alter their choosiness based on the availability of attractive males. There are many studies on sequential mate choice within species, but few have asked whether females apply these strategies to interactions between species and how these strategies may affect hybridization. We tested how previous interactions with conspecific and heterospecific males affect mate preference and sexual isolation in two threespine stickleback species (benthics and limnetics: Gasterosteus spp.). Consistent with previous work, we found that within species, stickleback females gauge male attractiveness relative to previously encountered males. If females extend these decision rules between species, we predicted that previous interactions with conspecifics should make heterospecifics less attractive, whereas interactions with heterospecifics should make conspecifics more attractive. However, females found heterospecifics less attractive after prior experience, largely independent of the species of male first encountered. Thus, sequential mate choice strategies are used within but not between species in sticklebacks. Further, learning from prior courtship interactions acts to enhance existing sexual isolation between species.     

Unilateral Mate Choice Causes Bilateral Behavioral Isolation Between Two Closely Related Phytophagous Ladybird Beetles (Coleoptera: Coccinellidae: Epilachninae)

Behavioral isolation between two sympatric and closely related phytophagous ladybird beetles, Henosepilachna vigintioctomaculata and Henosepilachana pustulosa, was studied in laboratory. Mating behavior was investigated using the male‐choice test. Males of H. vigintioctomaculata preferred conspecific females to heterospecific females as mates, whereas males of H. pustulosa did not show such preference. Females of H. vigintioctomaculata appeared more fastidious at mating than females of H. pustulosa, irrespective of the species of males. Males of neither species showed tenaciousness following rejection by females. The choice of conspecifics by H. vigintioctomaculata males and a difference in the intensity of rejection between H. vigintioctomaculata females (strong) and H. pustulosa females (weak) result in positive behavioral isolation in both directions of heterospecific matings. Our results thus indicated that positive unilateral mate choice yields bilateral behavioral isolation between these two species.