Does female control and male mating system predict courtship investment and mating outcomes? A comparative study in five widow spider species (genus Latrodectus) tested under similar laboratory conditions.

BACKGROUND: Male courtship investment may evolve in response to the male's expectation of future mating opportunities or the degree of female control during mating interactions. We used a comparative approach to test this hypotheses by assessing the courtship and mating behaviors of five widow spider species (genus Latrodectus) under common laboratory conditions. We predicted male investment in courtship would be higher in species where males mate only once because of high cannibalism rates (monogyny, L. geometricus, L. hasselti, L. mirabilis), compared to species with rare cannibalism (L. mactans, L. hesperus) in which males should reserve energy for future mating opportunities. Increased male investment, measured as courtship duration, might also evolve with increased female control over mating outcomes if females prefer longer courtships. We tested this by assessing the frequency of copulations, timing of sexual cannibalism, and the degree of female-biased size dimorphism, which is expected to be negatively correlated with the energetic cost of rebuffing male mating attempts. RESULTS: Copulation frequency was consistently lower in species with extreme female-skewed size dimorphism, and where sexual cannibalism was more prevalent, suggesting the importance of female control for mating outcomes. We confirmed significant interspecific variation in average courtship duration, but contrary to predictions, it was not predicted by male mating system, and there was no consistent link between courtship duration and sexual size dimorphism. CONCLUSION: We show that the degree of sexual dimorphism is not only correlated with sexual cannibalism, but also with mating success since restriction of male copulation frequency by female Latrodectus affects paternity. However, predictions about male mating system or female control affecting courtship duration were not supported. We propose that the form of female control over mating and cannibalism, and male responses, might be more informative for understanding the evolution of courtship duration. For example, male tactics to avoid female aggression may drive lower courtship duration in species like L. mirabilis. Nonetheless, our results differ from inferences based on published studies of each species in isolation, illuminating the need for standardized data collection for behavioural comparative studies.

The protamines of the noble false widow spider Steatoda nobilis provide an example of liquid-liquid phase separation chromatin transitions during spermiogenesis.

While there is extensive information about sperm nuclear basic proteins (SNBP) in vertebrates, there is very little information about Arthropoda by comparison. This paper aims to contribute to filling this gap by analyzing these proteins in the sperm of the noble false widow spider Steatoda nobilis (Order Araneae, Family Theridiidae). To this end, we have developed a protein extraction method that allows the extraction of cysteine-containing protamines suitable for the preparation and analysis of SNBPs from samples where the amount of starting tissue material is limited. We carried out top-down mass spectrometry sequencing and molecular phylogenetic analyses to characterize the protamines of S. nobilis and other spiders. We also used electron microscopy to analyze the chromatin organization of the sperm, and we found it to exhibit liquid-liquid phase spinodal decomposition during the late stages of spermiogenesis. These studies further our knowledge of the distribution of SNBPs within the animal kingdom and provide additional support for a proposed evolutionary origin of many protamines from a histone H1 (H5) replication-independent precursor.

Pre-copulatory choices drive post-copulatory decisions: mechanisms of female control shift across different life stages.

BACKGROUND: The 'wallflower' hypothesis proposes females mate indiscriminately to avoid reproductive delays. Post-copulatory mechanisms may then allow 'trading up', favouring paternity of future mates. We tested links between pre- and post-copulatory choice in Latrodectus geometricus female spiders paired sequentially with two males. These females copulate as adults or as subadults and store sperm in paired spermathecae. Choosy adults have a higher risk of delays to reproduction than subadults. RESULTS: We predicted low pre-copulatory, but high post-copulatory choice at first matings for adults and the opposite for subadults. At second matings, we expected all females would prefer males superior to their first. We found all females mated indiscriminately at their first pairing, but in contrast to subadults, adults usually allowed only a single insertion (leaving one of their paired spermatheca empty); a mechanism of post-copulatory choosiness. Adult-mated females were more likely to remate than subadult-mated females when they became adults, showing a preference for larger males, while subadult-mated females tended to prefer males of greater size-corrected mass. CONCLUSIONS: Our results show that the 'wallflower' effect and 'trading up' tactics can be utilized at different life stages, allowing females to employ choice even if rejecting males is costly.

Female control of a novel form of cannibalism during copulation in a South American widow spider.

Sexual cannibalism is an extreme form of sexual conflict that may have broad evolutionary effects on mating behaviour. Latrodectus spiders (∼30 "widow" species) could enable comparative tests of the evolution of sexual cannibalism, but most species are poorly-studied. Here we describe the mating and remating behaviour of a species endemic to South America, L. mirabilis, with a focus on the occurrence and consequences of sexual cannibalism. Mating and cannibalistic behaviours were novel relative to other Latrodectus species. Cannibalism started during copulation when females grasped the male's legs with their chelicerae, and continued as females pulled the male's abdomen onto their chelicerae, without interrupting copulation. Cannibalism was initiated by females, and not facilitated by males as was observed in other Latrodectus species. Females frequently remated however, so cannibalized males might lose paternity to future rivals. We report high rates of cannibalism, with 70 % of males killed by females during their first mating, and 85 % killed by previously-mated females. We discuss our novel findings in the context of previous observations in other Latrodectus species, proposing that foundational studies in a wider range of species are necessary to support comparative tests about the evolution of sexual cannibalism within this model taxon.

Immature mating as a tactic of polygynous male western widow spiders.

In polygynous species, males typically mate with more than one female, and male investment in any one mating should decrease if securing that mating reduces future mating opportunities. In contrast, monogynous males mate only once, so they should invest maximally in that single mating. In two monogynous, sexually cannibalistic Latrodectus spider species (L. geometricus, L. hasselti), males can mate and fertilize immature females. This recently described tactic is time-consuming because males must wait days for females to be capable of mating. It is also risky since immature females approached too early may kill males outright. However, if males typically find only one female in their lifetime, increasing the opportunity to mate may be worth these costs. We investigated whether this tactic is also practiced by a polygynous congener, L. hesperus, in which males typically mate more than once and may avoid time-consuming, risky encounters. In laboratory trials, we showed that males copulate with immature females. Moreover, males mounted immatures more rapidly, copulated for longer, and fathered more offspring than males that mated adults females. We concluded that monogyny is not a necessary condition for immature mating to be favored as an alternative reproductive tactic and suggest that it may be common in other spider taxa.

Neutral fitness outcomes contradict inferences of sexual 'coercion' derived from male's damaging mating tactic in a widow spider.

Sexual conflict over mating frequency has driven the evolution of morphological and behavioural traits across taxa. Interactions may be termed 'coercive' and assumed to arise from conflict when male mating behaviours cause physical injury to females and females appear to resist injurious matings.However, coercion per se occurs only if the behaviour reduces female fitness; and such outcomes are rarely measured. Here we show that a damaging mating tactic, apparently adaptive for males, is not coercive for females. Adult male Latrodectus spiders mate with immature females after tearing the exoskeleton covering the female's recently-developed reproductive tract, which can cause haemolymph bleeding. We show that, relative to pairings with adult females, males use reduced courtship displays when approaching immature females, which in some cases respond with elevated deterrent behavioural responses. Nevertheless, we found no reproductive cost for immature-mated females in terms of longevity, fertility or fecundity. Moreover, most immature-mated females did not produce sex pheromones as adults, so did not seek additional matings. Thus, despite the appearance of conflict there is no evidence that immature-mating is coercive. These results show it is critical to measure fitness outcomes, in addition to behavioural responses, to test for coercion.

Chemical communication in Schizocosa malitiosa: evidence of a female contact sex pheromone and persistence in the field.

Mate finding in many spider species is mediated, at least in part, by chemical cues. Although few have been characterized, most spider sex pheromones seem to be associated with the silk threads of adult females, attracting and/or triggering sexual behaviors in males. Schizocosa malitiosa (Araneae: Lycosidae) is a wolf spider common in dry environments in Southern Uruguay. Here, we report evidence for the occurrence of a female sex pheromone in the silk of virgin S. malitiosa females. The pheromone elicits typical courtship displays by conspecific males (palpal drumming and leg shaking), when it is applied to an artificial substrate. We also showed that this pheromone is quickly inactivated under field conditions, possibly due to the effect of dew, and that it is readily extracted from the silk by water, which renders the silk threads inactive. Preliminary chemical analyses by mass spectrometry suggest that the pheromone is a low molecular weight, highly oxygenated polar compound, present in a high concentration only in older virgin females. Quick inactivation in the field of the pheromone would allow males to discriminate between recent and old cues, thus facilitating mate searching and sexual meeting, by preventing males from following old cues for wandering females that could be distant in space and/or time.

Behavioural evidence of male volatile pheromones in the sex-role reversed wolf spiders Allocosa brasiliensis and Allocosa alticeps.

The use of chemical signals in a sexual context is widespread in the animal kingdom. Most studies in spiders report the use of female pheromones that attract potential sexual partners. Allocosa brasiliensis and Allocosa alticeps are two burrowing wolf spiders that show sex-role reversal. Females locate male burrows and initiate courtship before males perform any detectable visual or vibratory signal. So, females of these species would be detecting chemical or mechanical cues left by males. Our objective was to explore the potential for male pheromones to play a role in mate detection in A. brasiliensis and A. alticeps. We designed two experiments. In Experiment 1, we tested the occurrence of male contact pheromones by evaluating female courtship when exposed to empty burrows constructed by males or females (control). In Experiment 2, we tested the existence of male volatile pheromones by evaluating female behaviour when exposed to artificial burrows connected to tubes containing males, females or empty tubes (control). Our results suggest the occurrence of male volatile pheromones that trigger female courtship in both Allocosa species. The sex-role reversal postulated for these wolf spiders could be driving the consequent reversal in typical pheromone-emitter and detector roles expected for spiders.

Mating system in the tarantula spider Eupalaestrus weijenberghi (Thorell, 1894): evidences of monandry and polygyny.

The road tarantula Eupalaestrus weijenberghi shows a strongly female-biased sex ratio since adult females live several years while adult males live only for 2 months. In this scenario selective males could be expected. However, several factors such as the rates of reproduction of each sex, degree of sexual selectivity and synchronicity of female receptiveness determine the operational sexual ratio and mating system of the species. Our objective was to determine the mating rates and mating tactics for females and males of E. weijenberghi and their variation throughout the reproductive period. Four hundred sexual encounters among 20 females and 20 males in all possible pair-wise combinations were carried out during 29 days, a brief but intense experimental period, as it also occurs in the field. Mating success differed strongly between sexes. Females mated once: five females mated at the first attempt, eight initially rejected males and copulated in subsequent attempts. Half of the males did not copulate and the others copulated 1-3 times. Mated females actively rejected males. Results indicate a mating system with monogamous females and polygamous males. Not all the females were receptive in every reproductive season. We suggest that female monogamy drastically affects the operational sex ratio, since several females were unreceptive after a single copulation, directly diminishing the male potential reproductive rate. This is the first experimental approach to estimate tarantula mating systems, their determinants and the consequences of the strategies shown by each sex.