Female Mating Status Research Articles

Lethal Male Combat of Anastatus japonicus (Hymenoptera: Eupelmidae), an Egg Parasitoid of Lepidopterous and Hemipterous Pests.

Aggressive interactions between males are common when victors gain increased mating success but can result in severe injury or death for the defeated. Anastatus japonicus (Hymenoptera: Eupelmidae) is a solitary egg parasitoid of hemipteran and lepidopteran species. Here, we investigated lethal interactions between A. japonicus males and analyzed aggression behavior scaled with the male condition, number of competitors, number of presented females, and female mating status. The intensity of A. japonicus male aggressiveness increased with the number of competitors and the number of presented females but was not affected by male relatedness. The intensity and frequency of A. japonicus male aggressive interactions were greater when males competed for virgin females compared to mated females. Larger A. japonicus males won more contests than smaller males and honey-fed males defeated starved ones, where defeated males suffered the loss of appendages and/or died. Fighting appeared to be the norm in all-male groupings, regardless of the presence of females. A number of biological and ecological factors are deducted that likely contribute to the evolution of male aggressive behavior in A. japonicus.

Biotic factors as key determinants for ovarian and oothecal developmental plasticity of a tortoise beetle

Ovarian development in r-selected species is a highly dynamic process widely studied in various insect groups. An array of biotic and abiotic factors may influence it. So, the present investigation was to evaluate the impact of a female's age and mating status on the ovarian development, ootheca formation, body colour polymorphism, and fat content of the tortoise beetle, Aspidomorpha miliaris (Fabricius). Females of a certain age and mating status were dissected to demonstrate changes in their ovarioles, lateral oviduct, accessory gland, elytral colour, fat body content, and body size. It was predicted that age or mating status would not affect the ovarian parameters like length and width of ovarioles, lateral oviducts, accessory glands, body size, fat body content, and elytral colour. However, the ootheca-forming modifications would be initiated by ageing and mating. The current study demonstrated the substantial effect of age and mating status on the growth of the accessory glands and ovarioles. On the other hand, fat body contents declined comparatively in multiply-mated females. Besides this, the beetle exhibits elytral colour polymorphism till sexual maturity. An accessory gland was exclusively detected in multiply-mated females. This discovery opens opportunities for further investigation into the precise function of the gland, which is likely involved in the formation of ootheca.

Sperm depletion and mate choice of the crab spider, Ebrechtella tricuspidata (Fahricius)

Abstract Mounting evidence suggests that male sperm may be gradually depleted after consecutive matings, resulting in the decline of female reproductive output in insect species. It is predicted that females may employ adaptive strategies to avoid the sperm-depleted males, such as mating multiply with different males and/or discriminating against previously mated males (MMs). Similarly, males may exhibit adaptive behaviors toward females varying in mating status. However, in spiders with males lacking primary copulatory organs and their pedipalps modified to transfer sperm, there are few studies on male mating potential and previous mating experience on their subsequent mate choice. In this study, we used a polyandrous crab spider, Ebrechtella tricuspidata, a sit-and-wait predator with female-biased sex ratio as a model system to ascertain whether 1) male mating experiences influence female reproductive fitness; 2) females respond differently to males varying in mating status; and 3) males respond differently to females varying in mating status. Our results showed that female fecundity was independent of male mating experiences, but female fertility markedly declined with the increase of male previous mating experiences in the first eggsac. Counter to our predictions, females preferred to choose and mate with the larger males, regardless of their mating status. In contrast, male mating status influenced their mating preference toward females. Virgin males did not show any preference between virgin and mated females; however, MMs were more likely to prefer the virgin females over the mated ones. Overall, our results indicated that female choice depended primarily on male relative size rather than male mating status, whereas male choice depended on female mating status. Depending on the investigation of reproductive dynamics and mate choice in E. tricuspidata, the study will provide valuable insights into the adaptive behaviors of both males and females exhibiting toward mates varying in mating status.

Fake news? The impact of information mismatch on mating behaviour.

Multiple cues are often used for mate choice in complex environments, potentially entailing mismatches in the information conveyed by different sources. We address the consequences of this information mismatch for receivers using the spider mite Tetranychus urticae, in which virgin females are highly valuable mates compared to mated females, given first male sperm precedence. Accordingly, males are known to prefer virgins and distinguish them using cues from the females themselves and that they leave on the substrate. Whereas cues from females are highly reliable, those left on the substrate may not reflect the real female mating status if females move and/or mate. Here, we tested the consequences of such mismatch by exposing males to mated or virgin females on patches previously impregnated with cues deposited by females of either mating status. Male mating attempts were solely affected by substrate cues while female acceptance and the number of mating events were independently affected by both cues. Copulation duration, in contrast, depended mainly on the mating status of the female, with the number of copulations and the total time spent mating being intermediate in environments with mismatched information. We also show that males incur mating costs, reflected in reduced survival in environments with virgin cues. These results suggest that substrate cues left by females are instrumental for males to find their mates. However, in environments with mismatched information, males may pay survival costs without the associated benefit of mating with virgins, or they may lose opportunities to mate with virgins by responding to substrate cues from mated females. The benefit of using multiple cues will then hinge upon the frequency of information mismatch, which itself should vary with the dynamics of populations.

Unraveling the Significance of Draglines: Female Sexual Signalization in the Nursery-Web Spider, Pisaura mirabilis.

Chemical signals used by animals to attract the opposite sex are well known in insects, but heavily understudied in spiders. We investigated the role of chemical signals in female draglines in a gift-giving spider, Pisaura mirabilis, using combined data from behavioral tests and high-performance liquid chromatography (HPLC). We also investigated whether the quality of sexual signalization is influenced by crucial factors, such as female spider ontogeny, nutritional status, and mating status. We found that draglines of adult (versus subadult) and hungry (versus fed) females stimulated male motivation to produce nuptial gift, and highly sexually excited males invested more silk in gift production than less sexually excited males. Unexpectedly, chemical signals of eggsac-carrying females were similarly sexually attractive to draglines of adult females not carrying eggsac. HPLC identified significant chemical differences in female draglines, but these differences did not always correspond to male behavior. The integration of behavioral and chemical approaches is required to better understand animal behavior in future research.

Morphological changes in female reproductive organs in the African monarch butterfly, host to a male-killing Spiroplasma.

Sexual selection and conflicts within and between sexes promote morphological diversity of reproductive traits within species. Variation in the morphology of diagnostic reproductive characters within species offer an excellent opportunity to study these evolutionary processes as drivers of species diversification. The African monarch, Danaus chrysippus (Linnaeus, 1758), is widespread across Africa. The species is polytypic, with the respective geographical ranges of the four colour morphs only overlapping in East Africa. Furthermore, some of the populations host an endosymbiotic bacterium, Spiroplasma, which induces son-killing and distorts the local host population sex-ratio, creating sexual conflicts between the females seeking to optimize their fecundity and the limited mating capacity of the rare males. We dissected females from Kenya, Rwanda and South Africa, where Spiroplasma vary in presence and prevalence (high, variable and absent, respectively), and conducted microscopy imaging of their reproductive organs. We then characterized the effect of population, female body size, and female mating status, on the size and shape of different genitalia characters of the D. chrysippus female butterflies. We showed that although the general morphology of the organs is conserved in D. chrysippus, female genitalia vary in size and shape between and within populations. The virgin females have smaller organs, while the same organs were expanded in mated females. Females from highly female-biased populations, where the male-killing Spiroplasma is prevalent, also have a larger area of their corpus bursae covered with signa structures. However, this pattern occurs because a larger proportion of the females remains virgin in the female-biased populations rather than because of male depletion due to the symbiont, as males from sex-ratio distorted populations did not produce significantly smaller nutritious spermatophores.

Female mating status has been overlooked in mate choice research: a comment on Richardson and Zuk.

Female mating status has been overlooked in mate choice research: a comment on Richardson and Zuk.

A Neural Circuit Controlling Virgin Female Aggression Induced by Mating-related Cues in Drosophila.

Females increase aggression for mating opportunities and for acquiring reproductive resources. Although the close relationship between female aggression and mating status is widely appreciated, whether and how female aggression is regulated by mating-related cues remains poorly understood. Here we report an interesting observation that Drosophila virgin females initiate high-frequency attacks toward mated females. We identify 11-cis-vaccenyl acetate (cVA), a male-derived pheromone transferred to females during mating, which promotes virgin female aggression. We subsequently reveal a cVA-responsive neural circuit consisting of four orders of neurons, including Or67d, DA1, aSP-g, and pC1 neurons, that mediate cVA-induced virgin female aggression. We also determine that aSP-g neurons release acetylcholine (ACh) to excite pC1 neurons via the nicotinic ACh receptor nAChRα7. Together, beyond revealing cVA as a mating-related inducer of virgin female aggression, our results identify a neural circuit linking the chemosensory perception of mating-related cues to aggressive behavior in Drosophila females.

The effect of individual state on the strength of mate choice in females and males.

Animals are thought to gain significant fitness benefits from choosing high-quality or compatible mates. However, there is large within-species variation in how choosy individuals are during mating. This may be because the costs and benefits of being choosy vary according to an individual's state. To test this, I systematically searched for published data relating the strength of animal mate choice in both sexes to individual age, attractiveness, body size, physical condition, mating status, and parasite load. I performed a meta-analysis of 108 studies and 78 animal species to quantify how the strength of mate choice varies according to individual state. In line with the predictions of sexual selection theory, I find that females are significantly choosier when they are large and have a low parasite load, thus supporting the premise that the expression of female mate choice is dependent on the costs and benefits of being choosy. However, female choice was not influenced by female age, attractiveness, physical condition, or mating status. Attractive males were significantly choosier than unattractive males, but male mate choice was not influenced by male age, body size, physical condition, mating status, or parasite load. However, this dataset was limited by a small sample size, and the overall correlation between individual state and the strength of mate choice was similar for both sexes. Nevertheless, in both males and females individual state explained only a small amount of variation in the strength of mate choice.

Unlike a virgin: a meta-analytical review of female mating status in studies of female mate choice

AbstractStudies of female mate choice commonly use virgin females as test subjects, either to control for the effects of mating or because virgin females are presumed to be more responsive to mating cues. Theory predicts that virgin females will be less choosy because they risk dying without mating. Moreover, in many species, females spend more of their lives mated than as virgins. Thus, the exclusive use of virgin females in studies of female mate choice may underestimate the strength or direction of female choice and fail to reflect natural mating decisions. We conducted a systematic meta-analysis of female mate choice studies focusing on three scenarios in which female choice might differ in virgin and mated females: reproductive isolation, inbreeding avoidance, and sexually transmitted disease. Using only virgin females was common (53% of 303 studies). In addition, 38% of studies lacked information on female mating history. Contrary to predictions, we found no evidence that virgin females were less choosy than mated females. Nevertheless, excluding mated females from studies of female mate choice leaves an important gap in our understanding of the role of female preferences in evolution. We therefore encourage future studies of female mate choice to consider the natural context of mate choice and include mated females as test subjects when relevant.

Functional transcriptome analyses of Drosophila suzukii midgut reveal mating-dependent reproductive plasticity in females

BackgroundInsect females undergo a huge transition in energy homeostasis after mating to compensate for nutrient investment during reproduction. To manage with this shift in metabolism, mated females experience extensive morphological, behavioral and physiological changes, including increased food intake and altered digestive processes. However, the mechanisms by which the digestive system responds to mating in females remain barely characterized. Here we performed transcriptomic analysis of the main digestive organ, the midgut, to investigate how gene expression varies with female mating status in Drosophila suzukii, a destructive and invasive soft fruit pest.ResultsWe sequenced 15,275 unique genes with an average length of 1,467 bp. In total, 652 differentially expressed genes (DEGs) were detected between virgin and mated D. suzukii female midgut libraries. The DEGs were functionally annotated utilizing the GO and KEGG pathway annotation methods. Our results showed that the major GO terms associated with the DEGs from the virgin versus mated female midgut were largely appointed to the metabolic process, response to stimulus and immune system process. We obtained a mass of protein and lipid metabolism genes which were up-regulated and carbohydrate metabolism and immune-related genes which were down-regulated at different time points after mating in female midgut by qRT-PCR. These changes in metabolism and immunity may help supply the female with the nutrients and energy required to sustain egg production.ConclusionOur study characterizes the transcriptional mechanisms driven by mating in the D. suzukii female midgut. Identification and characterization of the DEGs between virgin and mated females midgut will not only be crucial to better understand molecular research related to intestine plasticity during reproduction, but may also provide abundant target genes for the development of effective and ecofriendly pest control strategies against this economically important species.

Single-Pair and Small-Group Crosses of Aedes aegypti.

Aedes aegypti is an emerging model insect species used in genetics studies because of its ease of laboratory rearing, desiccation-resistant eggs, expanding genetic toolkit, and high-quality reference genome. Here, we describe procedures to isolate and sex virgin female and male mosquitoes and establish successful mating crosses. We also detail how to blood feed mosquitoes from these crosses, isolate individual or small groups of females for egg laying, condition these eggs for storage and hatching, and verify female mating status.

Morphology and mating behaviour in the millipede Megaphyllum unilineatum (C.L. Koch, 1838) (Myriapoda, Diplopoda, Julida) under laboratory conditions

Abstract Although morphological variation may have an effect on behaviour, there are only a few studies on julid millipedes in which the influence of the variability of some morphological traits on mating success has been explored. Hence, objectives of this study were to investigate mating behaviour in laboratory conditions and identify traits that could possibly be the target of pre-copulatory selection in the julid species Megaphyllum unilineatum. Behavioural sequences were quantified in three types of tests: a mating arena test, a female choice test, and a male choice test. Although the number of contacts with the first chosen partner (from the mating arena test) was greater than with newly offered individuals in choice tests, values of the sexual selection coefficient did not statistically confirm this preference. In addition, analyses of linear measurements (trunk height and width, length of the whole body, antennae, walking legs, and gonopod flagella) in individuals of different mating status were also conducted, as well as geometric morphometric analyses of size and shape of the antennae, heads, walking legs, and gonopod promeres and opisthomeres in such individuals. Antennal length and shape, head shape, and the walking legs shape, differed significantly, depending on the mating status of females. In males of different mating status, statistical significance was established only in the promere centroid size. The differences in certain behavioural sequences in M. unilineatum are similar to those previously reported in M. bosniense, while such similarity is not detected with respect to morphological variation in the mentioned species.

Male mating success evolves in response to increased levels of male-male competition.

Male-biased operational sex ratios can increase male-male competition and can potentially select for both increased pre- and postcopulatory male success. In the present study, using populations of Drosophila melanogaster evolved under male-biased (M) or female-biased (F) sex ratios, we asked whether (a) male mating success can evolve, (b) males are better at mating females that they have coevolved with, (c) males mating success is affected by female mating status, and (d) male mating success is correlated with their courtship effort. We directly competed M and F males for mating with (a) virgin ancestral (common) females, (b) virgin females from the M and F populations, and (c) singly mated females from the M and F populations. We also assessed the courtship frequency of the males when paired with mated M or F females. Our results show that M males, evolving under an increased level of male-male competition, have higher mating success than F males irrespective of the female evolutionary history. However, the difference in mating success is more pronounced if the females had mated before. M males also have a higher courtship frequency than F males, but we did not find any correlation between mating success and courtship frequency.

Factors Influencing the Efficacy of Novel Attract-and-Kill (ACTTRA SWD) Formulations Against Drosophila suzukii.

In the continental United States, the invasive spotted-wing drosophila (SWD), Drosophila suzukii Matsumura, has become a primary pest of multiple stone and soft-skinned fruits. A new innovative adjuvant formulation, ACTTRA SWD, mixed with a suitable insecticide, constitutes a novel attract-and-kill tactic to manage D. suzukii in fruit crops. We hypothesized that background odors present in crop fields, particularly odors from host fruits, negatively affect the effectiveness of this attract-and-kill formulation, as odors from these sources can compete for insect attraction. Additionally, we evaluated the influence of adult D. suzukii sex and physiological status (age and mating status), and fruit ripeness on its response to the ACTTRA SWD formulation. For this, we used two-choice bioassays to test the response of adult D. suzukii to three ACTTRA SWD formulations (named OR1, TD, and HOOK SWD) in the presence and absence of host fruits (blueberries, raspberries, blackberries, and strawberries). Odors from raspberries were significantly more attractive than those from the TD formulation mixed with spinosad (Entrust). For the HOOK SWD formulation and OR1+Entrust formulation, odors from all the fruit types tested were significantly more attractive than the adjuvants. Compared with females, male D. suzukii were more attracted to the TD formulation over the blueberry fruits. Additionally, age and female mating status but not fruit ripeness influenced D. suzukii attraction to both OR1 and TD formulations. The results from this study indicate that D. suzukii physiological status and host fruit availability impact the efficacy of new attract-and-kill adjuvants such as ACTTRA SWD.

Mating behaviour, mate choice and female resistance in the bean flower thrips (Megalurothrips sjostedti)

Many species of thrips (Thysanoptera) in the family Thripidae form mating aggregations, but the adaptive significance of these aggregations and the extent of male and female mate choice is poorly understood. We studied the mating behaviour of the bean flower thrips Megalurothrips sjostedti (Trybom) (Thysanoptera: Thripidae), which forms male aggregations and occurs across sub-Saharan Africa. We tested whether males choose mates by female age or mating status. No-choice mating bioassays with one male and one female were used to simulate the way males usually encounter only one female at a time in aggregations in the field. Virgin females violently resisted mating attempts by males, but we found no compelling evidence to establish whether this was indiscriminate or was screening suitable males. Younger males (1–2 days old) did not discriminate females by age (1–2 or 7–10 days old), but older males (7–10 days old) avoided mating with older females. Any male choice by female mating status (virgin or mated) was weak or absent. The mating behaviour of M. sjostedti shows broad similarities with that of other thrips species that form aggregations, but also shows some distinct and novel differences, which can help our understanding of the adaptive significance of aggregations.

Mechanisms driving component Allee effects during invasions: using a biological control agent as model invader

1. A demographic Allee effect refers to increasing per capita population growth with increasing abundance. It arises from component Allee effects, which exist when some component of individual fitness has a positive relationship with density. Newly arrived populations tend to be small, and their establishment success is influenced by various factors, including demographic Allee effects. Identifying mechanisms driving the Allee effect is of relevance for understanding establishment failure, developing strategies for improving biological control and conservation, and for managing biological invasions.2. We utilised an invasive plant biocontrol agent, Neolema ogloblini, as model to study Allee effects experienced by invading populations. We investigated mating failure and predator satiation as two component Allee effects that could potentially drive a demographic Allee effect in N. ogloblini populations.3. We released unmated adults onto isolated host patches using five release sizes and evaluated the mating status of females after 3 weeks in the field. The probability of being mated increased as number of males recovered increased, suggesting the presence of a mating‐failure component Allee effect in very small populations of N. ogloblini. Exclosures were used to evaluate survival of immatures of N. ogloblini developing in small group sizes, either in the presence or absence of generalist predators. Results revealed high levels of predation, but a positive relationship between larval survival and group size in the presence of generalist predators could not be verified.4. Our study identified relevant life‐history traits contributing to establishment failure in N. ogloblini and demonstrates the value of biocontrol agents as models to study mechanisms of species invasion.

Males of a Neotropical spider adjust prey‐gift construction but not mate search in response to sperm competition

AbstractIn scramble competition mating systems, males do not guard females and females usually mate polyandrously, leading to high levels of sperm competition. Mathematical models predict that males facing sperm competition can either decrease or increase their reproductive effort. To test how male reproductive responses vary according to sperm competition risk, we ran two experiments using the polyandrous gift‐giving spiderParatrechalea ornata. First, we tested whether males prefer substrates containing silk of unmated females than substrates containing silk of once‐mated females, competitor males or without silk. Males spent more time on substrates containing silk of females but showed no preference for unmated females, which indicates that males may optimize mate search by prioritizing substrates containing cues of potential mating partners. The lack of preference for unmated females suggests that males cannot detect female mating status from silk or that the costs of searching preferentially for unmated females are high in the field. Second, we tested whether males adjust prey‐gift construction when facing sperm competition risk. Experimental groups (substrate containing silk of an unmated female and silk of another malevs. substrate containing only silk of an unmated female) did not differ in the frequency of gift construction, latency to gift construction and number of flies added to the gift. However, males facing sperm competition risk constructed gifts with less silk, suggesting the occurrence of a less evident form of male mate choice based on differential allocation of resources to prey‐gift. Additionally, given the trade‐off between investing in prey‐gifts quantity or quality, a lower silk investment should enable males to increase the number of gifts constructed and the number of courted females. Taken together, our results indicate that males detect female and male pheromones on substrate and adjust the construction of prey‐gifts in response to apparent sperm competition risk.

Female mating status affects male mating tactic expression in the wolf spider Rabidosa punctulata.

Males and females have conflicting interests on the frequency and outcomes of mating interactions. Males maximize their fitness by mating with as many females as possible, whereas choosy females often reduce receptivity following copulation. Alternative male mating tactics can be adaptive in their expression to a variety of mating contexts, including interactions with a relatively unreceptive mated female. Male Rabidosa punctulata wolf spiders can adopt distinctive mating tactics when interacting with a female, a complex courtship display, and/or a more coercive direct mount tactic that often involves grappling with females for copulation. In this study, we set up female mating treatments with initial trials and then paired mated and unmated females with males to observe both female remating frequencies and the male mating tactics used during the interactions. Males adopted different mating tactics depending on the mating status of the female they were paired with. Males were more likely to adopt a direct mount tactic with already-mated females and courtship with unmated females. Already-mated females were considerably less receptive to males during experimental trials, although they did remate 34% of the time, the majority of which were with males using a direct mount tactic. Whereas males adjusting to these contextual cues were able to gain more copulations, the observation of multiple mating in female R. punctulata introduces the potential for sperm competition. We discuss this sexual conflict in terms of the fitness consequences of these mating outcomes for both males and females.

Drosophila oocyte proteome composition covaries with female mating status

Oocyte composition can directly influence offspring fitness, particularly in oviparous species such as most insects, where it is the primary form of parental investment. Oocyte production is also energetically costly, dependent on female condition and responsive to external cues. Here, we investigated whether mating influences mature oocyte composition in Drosophila melanogaster using a quantitative proteomic approach. Our analyses robustly identified 4,485 oocyte proteins and revealed that stage-14 oocytes from mated females differed significantly in protein composition relative to oocytes from unmated females. Proteins forming a highly interconnected network enriched for translational machinery and transmembrane proteins were increased in oocytes from mated females, including calcium binding and transport proteins. This mating-induced modulation of oocyte maturation was also significantly associated with proteome changes that are known to be triggered by egg activation. We propose that these compositional changes are likely to have fitness consequences and adaptive implications given the importance of oocyte protein composition, rather than active gene expression, to the maternal-to-zygotic transition and early embryogenesis.