Fitness advantage from nuptial gifts in female fireflies

Abstract 1. In many insects, males provide nuptial gifts to females in the form of spermatophores, sperm-containing structures produced by male accessory glands.
2. The work reported here examined the influence of both spermatophore number and spermatophore size on female reproductive output in two related firefly beetles, Photinus ignitus and Ellychnia corrusca (Coleoptera: Lampyridae). Based on differences in adult diet, male spermatophores were predicted to increase female reproductive output to a greater extent in P. ignitus than in E. corrusca .
3. Female fecundity was significantly higher in triply mated females than in singly mated females in both species, with no difference between mating treatments in female lifespan or egg hatching success. No effects of second male spermatophore size on fecundity, lifespan, or egg hatching success were detected in either species.
4. These results suggest a direct fitness advantage from multiple mating for females in both species, although enhanced fecundity may be due either to allocation of spermatophore nutrients to eggs or to other substances transferred within the spermatophore acting as oviposition stimulants.     

Nuptial gifts and sexual selection in photinus fireflies

The phenomenon of nuptial gift transfer during mating occursacross a remarkably wide range of taxa, and such male donationsare likely to influence both pre-copulatory and post-copulatorysexual selection. This paper reviews what is known about nuptialgifts in Photinus fireflies (Coleoptera: Lampyridae), and discussesthe adaptive significance of spermatophores in firefly matingsystems. During copulation Photinus males transfer a spiral,gelatinous spermatophore to the female: sperm are released intothe female's spermatheca for storage, while the remainder ofthe spermatophore disintegrates within a specialized gland.Radiolabelling studies indicate that male-derived protein isused to help provision the female's developing oocytes, andmultiply-mated females show increased fecundity. As most Photinusadults do not feed, these studies suggest that females shouldcontinue to forage for matings to supplement their diminishinglarval reserves, even after they have gained sufficient spermto fertilize their eggs. Male spermatophore mass declines acrosssequential matings, and smaller spermatophores are associatedwith lower paternity success in situations where males competefor fertilizations. Declining spermatophore size across sequentialmatings may thus lead to diminishing reproductive returns forfirefly males. Taken together, these results suggest that seasonalchanges in nuptial gift availability may contribute to reversalsof traditional courtship roles, with male choice and female-femalecompetition occurring as spermatophore availability declines.     

Sperm storage mediated by cryptic female choice for nuptial gifts

Polyandrous females are expected to discriminate among males through postcopulatory cryptic mate choice. Yet, there is surprisingly little unequivocal evidence for female-mediated cryptic sperm choice. In species in which nuptial gifts facilitate mating, females may gain indirect benefits through preferential storage of sperm from gift-giving males if the gift signals male quality. We tested this hypothesis in the spider Pisaura mirabilis by quantifying the number of sperm stored in response to copulation with males with or without a nuptial gift, while experimentally controlling copulation duration. We further assessed the effect of gift presence and copulation duration on egg-hatching success in matings with uninterrupted copulations with gift-giving males. We show that females mated to gift-giving males stored more sperm and experienced 17% higher egg-hatching success, compared with those mated to no-gift males, despite matched copulation durations. Uninterrupted copulations resulted in both increased sperm storage and egg-hatching success. Our study confirms the prediction that the nuptial gift as a male signal is under positive sexual selection by females through cryptic sperm storage. In addition, the gift facilitates longer copulations and increased sperm transfer providing two different types of advantage to gift-giving in males.     

Multiple mating and lifetime reproductive output in female dobsonflies that receive nuptial gifts

In mating of the dobsonfly, Protohermes grandis (Thunberg), the male attaches the spermatophore externally to the female genitalia. The spermatophore includes a large gelatinous mass which the female detaches and feeds on after mating. While the female consumes this nuptial food gift, sperm is evacuated from the remaining portion of the spermatophore (sperm package) into her reproductive tract. Under laboratory conditions, mated females maintained receptivity throughout their lifetime, and they remated even on the day following copulation. A single insemination may supply enough sperm, as females mated only once deposited fertile eggs throughout life and, when dissected after death, all females had sperm in the spermatheca. There was a positive correlation between longevity and the number of matings. Lifetime fecundity also increased as mating multiplied. However, the size of eggs and hatchlings was not influenced by the number of matings. It seems that large spermatophore consumption by female P. grandis provides nutrients that increase fitness not in offspring quality, but in their quantity.     

Allometry and proximate mechanisms of sexual selection in photinus fireflies, and some other beetles

Ever since Darwin identified it as the force responsible forthe evolution of exaggerated male characters, sexual selectionhas been the focus of research aimed at understanding the mostbizarre and intriguing morphologies and behaviors in Nature.Two congeneric species in the firefly genus Photinus, P. pyralisand P. macdermotti, afford a unique opportunity to examine theinteraction between sexual and countervailing natural selectionthat act to shape the evolution of mating behavior and bodysize in closely related species with very different courtshipstrategies. Photinus pyralis males emit very bright flashesduring their extended patrolling flights and form ‘loveknots’ of competing suitors, while P. macdermotti males,whose courtships seldom exceed two competing males, produceweaker flashes during a shorter patrolling period. Possibly as a consequence of their scramble competition andlong flights, not only is there an extremely wide range of bodysizes in P. pyralis males, but they also exhibit wing allometricslopes greater than one, and lantern allometries less than one.In contrast, P. macdermotti males do not have allometric slopessignificantly different from one. Small males, when tested inan artificial scramble situation move faster than large males,an advantage in the intense competition that ensues once a femaleis located. Females answer several males in alternation andfail to respond to all male flashes, with the evident consequencethat love knots frequently develop around such fickle females.Allometric relationships in two non-luminescent beetle specieswith non-visual courtship protocols are compared. Visual, sexuallyselected characters showed positive allometric slopes, whilenon-visual characters showed isometry or negative slopes. Datapresented here support the existence of distinct patterns inmodality-specific sexual selection.     

Male courtship signals and female signal assessment in Photinus greeni fireflies

The evolutionary dynamic of courtship signaling systems is drivenby the interaction between male trait distributions and femalepreferences. This interaction is complex because females maychoose mates based on multiple components of male signals, andfemale preference functions may vary depending on mate availability,female reproductive state, and environmental conditions. InPhotinus fireflies (Coleoptera: Lampyridae), flying males emitbioluminescent flash signals to locate sedentary females, whichreply selectively to attractive male flash signals with theirown response flash. In this study, we first examined temporalvariation in the paired-pulse flash patterns produced by Photinusgreeni males in the field and found significant among-male variation(70% of total variation) in interpulse intervals (IPIs). Therewas no significant relationship between male IPI and spermatophoresize, suggesting that P. greeni male courtship signals do notprovide females with reliable indicators of male material resources.In laboratory playback experiments, we presented P. greeni femaleswith simulated flash signals to assess how IPI and pulse durationindependently affected the likelihood of female flash response.We also examined the effects of female body mass and time duringthe mating season on female preference functions, hypothesizingthat females would be less discriminating when they were heavier(more fecund) and when mate availability declined. We foundthat P. greeni females discriminated among signals within theirspecies' range based primarily on flash pattern IPI. Neitherthe time during the mating season nor female weight alteredfemale preference functions for IPI, although season did influencefemale response to pulse duration. These results reveal thatP. greeni females discriminate among conspecific males basedprimarily on male IPIs, the same signal character previouslyshown to be important for firefly species recognition. Fieldplayback experiments indicated that female responsiveness peakednear the average IPI given by males at different ambient temperatures,suggesting that fireflies exhibit temperature coupling similarto that seen in many acoustically signaling animals.     

Mating stimulates female feeding: testing the implications for the evolution of nuptial gifts

Nutritional benefits from nuptial gifts have been difficult to detect in some species, raising the question: what maintains nuptial feeding when gifts do not benefit females? The sensory trap hypothesis proposes that nuptial feeding may be explained by pre‐existing sensory responses that predispose females to ingest gifts. Recent studies have shown that male seminal proteins can induce a nonspecific increase in female feeding after mating, which may represent a sensory trap for nuptial feeding if it results in increased intake of post‐mating gifts. I tested these ideas using female beetles that ingest a spermatophore after mating. I show that males stimulate strongly increased female feeding post‐mating. However, there was little evidence for dose dependence in the feeding response that could allow males to stimulate feeding beyond the female optimum. Moreover, the post‐mating feeding response could not explain nuptial feeding: despite feeding more in general, newly mated females were less likely than nonmated females to ingest spermatophore gifts.     

Emerging issues in the evolution of animal nuptial gifts

Uniquely positioned at the intersection of sexual selection, nutritional ecology and life-history theory, nuptial gifts are widespread and diverse. Despite extensive empirical study, we still have only a rudimentary understanding of gift evolution because we lack a unified conceptual framework for considering these traits. In this opinion piece, we tackle several issues that we believe have substantively hindered progress in this area. Here, we: (i) present a comprehensive definition and classification scheme for nuptial gifts (including those transferred by simultaneous hermaphrodites), (ii) outline evolutionary predictions for different gift types, and (iii) highlight some research directions to help facilitate progress in this field.     

Paternity of offspring in multiply-mated,female crickets: the effect of nuptial food gifts and the advantage of mating first

The spermatophore transferred by male decorated crickets (Gryllodes sigillatus) includes a large gelatinous mass, the spermatophylax, that is consumed by the female after mating. This nuptial gift preoccupies the female while sperm are discharged from the remaining portion of the spermatophore, the sperm ampulla, into her reproductive tract. There is considerable variation in the mass of the spermatophylax, and about half of all males produce spermatophylaxes that are too small to ensure complete sperm transfer. We tested two hypotheses concerning the maintenance of this variation: (i) males trade-off investment in spermatophylaxes against copulation frequency; and (ii) males synthesize the largest spermatophylaxes of which they are physiologically capable. Males synthesizing large and small food gifts were permitted multiple mating opportunities with the same females, and allozyme markers were used to establish the paternity of offspring. There was a significant advantage to those males that mated first irrespective of gift size. This advantage probably arose, in part, because the sperm of first males would have had exclusive access to females'' eggs during the first 24 hours of oviposition, and underscores the benefits of matings with virgin females. The paternity of ''small-gift'' males increased with gift mass, but there was no such increase in ''large-gift'' males. This difference probably stems from the relationship between gift mass and sperm transfer: most of the gifts of the large-gift males would have been above the threshold needed to achieve complete inseminations, whereas those of small-gift males would have been below the threshold. Within mating-order positions, there was no significant difference in the paternity of large-gift and small-gift males, a result seemingly consistent with the ''trade-off'' hypothesis. However, there was no correlation between spermatophylax mass and male mating frequency, so that the mechanism by which small-gift males offset their fertilization disadvantage remains unknown.     

The energetics of trading nuptial gifts for copulations in katydids

During copulation, male Isophya kraussi transfer a large nuptial gift to females. In this study, we hypothesized that the energy content of spermatophores should meet the energy requirements of both body maintenance and egg production of females. We measured the field metabolic rate of male and female I. kraussi using the doubly labeled water method and the energy content of spermatophores and male bodies with microbomb calorimetry. The energy content of male nuptial gifts averaged 0.66+/-0.09 kJ, approximately 20% of the total body energy content of male I. kraussi (3.24+/-0.26 kJ). Field metabolic rates averaged 0.41+/-0.17 kJ d(-1) (n = 8) for males and 0.30+/-0.15 kJ d(-1) (n = 5) for female I. kraussi. Thus, the energy content of spermatophores exceeded the daily energy requirements of existence in male I. kraussi. A single nuptial gift provides for all energy requirements of females for 1 or 2 d, depending on their activity, egg production, and ambient temperature. Because the shortest known remating interval of female katydids varies between 1 and 3 d, female I. kraussi could theoretically exist exclusively on spermatophores to meet their nutritional requirements.     

Molecular dissection of nuptial gifts in divergent strains of Ostrinia moths

Seminal fluid proteins (SFPs) produced in the male accessory glands and ejaculatory duct are subject to strong sexual selection, often evolve rapidly and therefore may play a key role in reproductive isolation and species formation. However, little is known about reproductive proteins for species in which males transfer ejaculate to females using a spermatophore package. By combining RNA sequencing and proteomics, we characterize putative SFPs, identify proteins transferred in the male spermatophore and identify candidate genes contributing to a one‐way gametic incompatibility between Z and E strains of the European corn borer moth Ostrinia nubilalis. We find that the accessory glands and ejaculatory duct secrete over 200 highly expressed gene products, including peptidases, peptidase regulators and odourant‐binding proteins. A comparison between Ostrinia strains reveals that accessory gland and ejaculatory duct sequences with hormone degradation and peptidase activity are among the most extremely differentially expressed. However, most spermatophore peptides lack reproductive tissue bias or canonical secretory signal motifs and aproximately one‐quarter may be produced elsewhere before being sequestered by the male accessory glands during spermatophore production. In addition, most potential gene candidates for postmating reproductive isolation do not meet standard criteria for predicted SFPs and almost three‐quarters are novel, suggesting that both postmating sexual interactions and gametic isolation likely involve molecular products beyond traditionally recognized SFPs.     

Terminal investment in the gustatory appeal of nuptial food gifts in crickets

Investment in current versus future reproduction represents a prominent trade‐off in life‐history theory and is likely dependent on an individual's life expectancy. The terminal investment hypothesis posits that a reduction in residual reproductive value (i.e. potential for future offspring) will result in increased investment in current reproduction. We tested the hypothesis that male decorated crickets (Gryllodes sigillatus), when cued to their impending mortality, should increase their reproductive effort by altering the composition of their nuptial food gifts (i.e. spermatophylaxes) to increase their gustatory appeal to females. Using a repeated‐measures design, we analysed the amino acid composition of spermatophylaxes derived from males both before and after injection of either a saline control or a solution of heat‐killed bacteria. The latter, although nonpathogenic, represents an immune challenge that may signal an impending survival threat. One principal component explaining amino acid variation in spermatophylaxes, characterized by a high loading to histidine, was significantly lower in immune‐challenged versus control males. The relevance of this difference for the gustatory appeal of gifts to females was assessed by mapping spermatophylax composition onto a fitness surface derived in an earlier study identifying the amino acid composition of spermatophylaxes preferred by females. We found that immune‐challenged males maintained the level of attractiveness of their gifts post‐treatment, whereas control males produced significantly less attractive gifts post‐injection. These results are consistent with the hypothesis that cues of a survival‐threatening infection stimulate terminal investment in male decorated crickets with respect to the gustatory appeal of their nuptial food gifts.     

Sensory exploitation as an evolutionary origin to nuptial food gifts in insects

Nuptial food gifts given by males to females at mating are widespread in insects, but their evolutionary origin remains obscure. Such gifts may arise as a form of sensory trap that exploits the normal gustatory responses of females, favouring the selective retention of sperm of gift-giving males. I tested this hypothesis by offering foreign food gifts, synthesized by males of one cricket species, to females of three non-gift-giving species. Females provisioned with novel food gifts were 'fooled' into accepting more sperm than they otherwise would in the absence of a gift. These results support the hypothesis that nuptial food gifts and post-copulatory female mating preferences coevolve through a unique form of sensory exploitation.     

Unpredictable environments,nuptial gifts and the evolution of sexual size dimorphism in insects: an experiment

Many insects have a mating system where males transfer nutrients to females at mating, which are often referred to as ''nuptial gifts''. Among butterflies, some of the characteristic features of these species are polyandry (females mate multiple times), and relatively large male ejaculates. When males produce part of the resources used for offspring, the value of body size might then increase for males and decrease for females. The male/female size ratio is also observed to increase when the degree of polyandry and gift size increase. Butterfly species where gift-giving occurs are generally more variable in body size, suggesting that food quality/quantity fluctuates during juvenile stages. This will cause some males to have much to provide and some females to be in great need, and could be conducive to the evolution of a gift-giving mating system. In such a system, growing male and female juveniles should react differently to food shortage. Females should react by maturing at a smaller size since their own lack of reproductive resources can partly be compensated for by male contributions. Males have to pay the full cost of decreased reproduction if they mature at a small size, making it more important for males to keep on growing, even when growth is costly. An earlier experiment with the polyandrous and gift-giving butterfly, Pieris napi, supported this prediction. The pattern is expected to be absent or reversed for species with small nuptial gifts, where females do not benefit from mating repeatedly, and will thus be dependent on acquiring resources for reproduction on their own. To test this prediction, we report here on an experiment with the speckled wood butterfly, Pararge aegeria. We find that growth response correlates with mating system in the two above species, and we conclude that differences in environmental conditions between species may act as an important factor in the evolution of the mating system and sexual size dimorphism.     

Female preference for male courtship flashes in Photinus ignitus fireflies

In Photinus fireflies, males produce spontaneous bioluminescentcourtship flashes. Females preferentially respond to particularmale flashes with flashes of their own. This study exploredvariation in female flash responsiveness as a function of maleflash duration, female condition, lantern size, and lanterndistance, as well as the relationship between male characteristicsand spermatophore mass in Photinus ignitus fireflies. We determinedfemale preference by scoring female flash response to simulatedmale flashes and determined variation in overall female flashresponsiveness for laboratory-mated, laboratory-fed, and controlP. ignitus females. Flash duration, lantern size, and body masswere recorded for field-collected males. Males were then matedto determine spermatophore mass. Females exhibited greater preferencefor artificial flashes representing the upper range of conspecificmale flash duration and lantern size as well as flashes producedat a closer distance. Both laboratory-mated and laboratory-fedP. ignitus females showed lower overall responsiveness acrossall flash durations relative to control females that did notmate or feed in the laboratory. Male flash duration predicteda significant proportion of the variation in spermatophore massfor early-season males. These results suggest that female Photinusignitus may prefer long flashes in order to obtain the directbenefit of larger spermatophores and may adjust their overallflash responsiveness as the relative importance of this benefitvarieswith changing female condition.     

Multiple signals and male spacing affect female preference at cocktail parties in treefrogs

Effective acoustic communication in the face of intense conspecific background noise constitutes a constant sensory challenge in chorusing and colonial species. An evolutionary approach suggests that behavioural and environmental constraints in these species should have shaped signal design and signalling behaviour to enable communication in noisy conditions. This could be attained both through the use of multicomponent signals and through short-term adjustments in the spatial separation of calling males. We investigated these two hypotheses in a chorusing anuran, the hylid Hyla arborea, through a series of phonotaxis experiments conducted within a six-speaker arena in a high background noise situation, by presenting females with male calls containing either single or multiple attractive call components, and by modifying distances between speakers. We found that female ability to discriminate attractive calls increased when several attractive call components were available, providing novel evidence that the use of multicomponent signals enhances communication in complex acoustic conditions. Signal discrimination in females also improved with speaker separation, demonstrating that within natural choruses, spatial unmasking conditioned by male density and spatial separation probably improves female discrimination of competing males. Implications of these results for the accuracy of mate choice within choruses are discussed.