Mate choice among males is relatively understudied, despite recent evidence supporting its ubiquity. Theory predicts male mate choice in response to variation in female quality, and male mating strategies that limit polygyny. However empirical research investigating these connections, particularly under natural conditions, is generally lacking. Using the Western black widow spider, Latrodectus hesperus, I investigated male mate choice, male investment, and the advantage of mating with females that differ in their potential to deliver fitness benefits to males via higher fecundity and/or reduced risk of sperm competition. Males were found to be preferentially attracted to larger, high-diet virgin females over females that were smaller due to a lower diet, or having been previously mated, or both. Through a three-year field study on females I found that males likely benefit from selectivity, as smaller, low-diet females often failed to deposit any egg sacs throughout the breeding season, likely due to their shortened web-site tenure and/or lack of bodily resources. I investigated the costs and benefits to male genital breakage in L. hesperus, a strategy typically assumed to result in male sterility while providing paternity protection. I found that genital mutilation in L. hesperus did not cause sterility and that males were capable of inseminating multiple females, likely because of the comparatively low amount of genital damage and the likelihood of the efficient placement of genital fragments. Evidence from double mating trials supported the efficacy of broken genital fragments as plugs, but first male sperm precedence was often maintained in cases where male genital breakage failed or fragments were positioned incorrectly, which occurred frequently. Together, these laboratory and field experiments contribute to a more complete view of mate choice.

Mate choice among males is relatively understudied, despite recent evidence supporting its ubiquity. Theory predicts male mate choice in response to variation in female quality, and male mating strategies that limit polygyny. However empirical research investigating these connections, particularly under natural conditions, is generally lacking. Using the Western black widow spider, Latrodectus hesperus, I investigated male mate choice, male investment, and the advantage of mating with females that differ in their potential to deliver fitness benefits to males via higher fecundity and/or reduced risk of sperm competition. Males were found to be preferentially attracted to larger, high-diet virgin females over females that were smaller due to a lower diet, or having been previously mated, or both. Through a three-year field study on females I found that males likely benefit from selectivity, as smaller, low-diet females often failed to deposit any egg sacs throughout the breeding season, likely due to their shortened web-site tenure and/or lack of bodily resources. I investigated the costs and benefits to male genital breakage in L. hesperus, a strategy typically assumed to result in male sterility while providing paternity protection. I found that genital mutilation in L. hesperus did not cause sterility and that males were capable of inseminating multiple females, likely because of the comparatively low amount of genital damage and the likelihood of the efficient placement of genital fragments. Evidence from double mating trials supported the efficacy of broken genital fragments as plugs, but first male sperm precedence was often maintained in cases where male genital breakage failed or fragments were positioned incorrectly, which occurred frequently. Together, these laboratory and field experiments contribute to a more complete view of mate choice.

Postcopulatory sexual selection—sperm competition and cryptic female choice—has become a major area of research over the past 40 years. Within this field there are many outstanding questions at every level of analysis, from proximate to ultimate. The fitness consequences for both sexes in the period after copulation and before fertilization are considerable, but are obscured within the female reproductive tract. Our understanding of postcopulatory mechanisms is especially sparse in taxa other than birds and insects. Nearly nothing is known in reptiles except that multiple paternity is common and widespread, and often results from long-term sperm storage across breeding seasons. We present some of the very first data on the determinants of fertilization success in the context of sperm competition in reptiles, a group that accounts for 30% of terrestrial vertebrates. In the first chapter, "Asymmetric gametic isolation between two populations of red-sided garter snakes", we discuss the use of between-population crosses to reveal gametic isolation. The effect of population density and operational sex ratios on mating systems and the speciation process has fueled theoretical debate. We attempted to address these issues using two populations of red-sided garter snakes (Thamnophis sirtalis parietalis) from Manitoba, Canada. Our study populations differ markedly in their density mating aggregations, with a 10-fold difference between them. Using microsatellite markers for paternity analysis of litters produced from within and between population crosses. We found that the population with highest aggregation density, and presumably with the highest level of sexual conflict (i.e., when the evolutionary interests of the sexes differ) over mating, was also the population that exhibited homotypic sperm precedence. The less dense population showed a distinct postcopulatory male-size advantage. We also demonstrated that sperm stored within the female over hibernation can father 20-30% of offspring in a litter. In the second chapter, "Sperm competition and mate-order effects in red-sided garter snakes", we test whether females use mate-order effects to ensure that a larger (fitter) male will sire her offspring. Does that second male should have precedence in sperm competition? We tested for second-male precedence using singly-mated females that mated with a second male. Average proportion of paternity was shared equally among the first (P₁, i.e., proportion of offspring from a litter fathered by the first male to mate) and second males (P₂) to mate, and stored sperm (P[subscript ss]). This may be a case where last male precedence breaks down with more than two males. All females were spring virgins (they had not mated that spring, but may have stored sperm from fall matings); thus sperm stored presumably from fall matings is important in this system. As the interval between matings increased P₁ increased at the expense of P[subscript ss]. As the second male to mate's copulation duration increased, P₁ also increased at the expense of P₂. This last result may indicate female influence over sperm transfer during coerced matings. Copulatory plugs (CPs) are found in many taxa, but the functional significance is debated. Male garter snakes produce a gelatinous copulatory plug during mating that occludes the opening of the female reproductive tract for approximately two days. In chapter three, "Not just a chastity belt: the role of mating plugs in red-sided garter snakes revisited", we experimentally tested the role of the CPs. In snakes, sperm are produced in the testes and delivered through the ductus deferens, and the copulatory plug is thought to be produced by the sexual segment of the kidney and conveyed through the ureter. We manipulated the delivery of the two fluids separately by ligating the ducts. We confirmed that the CP is not formed in ureter-ligated males and that sperm leaks out immediately after copulation. The CP is analogous to a spermatophore. The protein matrix contains most of the sperm which are liberated as the plug dissolves within the female's vaginal pouch. One of the fundamental principles in sperm competition is that increased sperm numbers increase the odds of winning in competitions for fertilization success and males will adjust their ejaculate relative to competition and the quality of his mate. In chapter four, "Sperm depleted males and the unfortunate females who mate with them", we detect significant among-male variation in the number of sperm ejaculated, and that male mate-order reduces sperm numbers. Male sperm numbers drop significantly from one mating to the next, and this has implications for sperm competiveness, as Thamnophis sirtalis exhibits a disassociated reproductive tactic, in that sperm stores are produced outside the breeding season, and thus cannot be replenished after mating. Interestingly, however, the on average the mobility of the sperm increased for a male's second mating. Therefore, increased sperm quality may compensate for reduced numbers in a competitive context. Further, females increase their remating rate when mating with males that are unable to deliver sperm. In chapter five, "Sexual conflict during mating in red-sided garter snakes as evidenced by genital manipulation", we revisited the CP in the context of sexual conflict. Sex-differences in optimal copulation duration can be a source of conflict, as increased copulation duration may be advantageous for males as it delays female remating. Males of many species actively guard females to prevent them from remating, and in some cases males produce copulatory plugs to prevent remating. If precopulatory choice is limited at the time of her first mating, conflict may be especially onerous to a female. The size of the plug is influenced by the copulation duration. We experimentally tested the contribution of male and female control over copulation duration. We ablated the largest basal spine on the male's hemipene and found a reduction in copulation duration and an increase in the variation of plug mass. Further, we anesthetized the female's cloaca and found copulation duration increased, which suggests that males benefit from increased copulation duration while females actively try to reduce copulation duration. Therefore, sexual conflict is manifest in divergent copulation duration optima for males and females. / Graduation date: 2013