Anne Lizé

Kin recognition in Drosophila: the importance of ecology and gut microbiota

The animal gut commonly contains a large reservoir of symbiotic microbes. Although these microbes have obvious functions in digestion and immune defence, gut microbes can also affect behaviour. Here, we explore whether gut microbiota has a role in kin recognition. We assessed whether relatedness, familiarity and food eaten during development altered copulation investment in three species of Drosophila with diverse ecologies. We found that a monandrous species exhibited true kin recognition, whereas familiarity determined kin recognition in a species living in dense aggregations. Finally, in a food generalist species, food eaten during development masked kin recognition. The effect of food type on copulation duration, in addition to the removal of this effect via antibiotic treatment, suggests the influence of bacteria associated with the gut. Our results provide the first evidence that varied ecologically determined mechanisms of kin recognition occur in Drosophila, and that gut bacteria are likely to have a key role in these mechanisms.

Kin discrimination and altruism in the larvae of a solitary insect

Kin selection theory predicts altruism between related individuals, which requires the ability to recognize kin from non-kin. In insects, kin discrimination associated with altruistic behaviour is well-known in clonal and social species but in very few solitary insects. Here, we report that the solitary larvae of a non-social insect Aleochara bilineata Gyll. (Coleoptera; Staphylinidae) show kin discrimination and sibling-directed altruistic behaviour. Larvae superparasitize more frequently the hosts parasitized by non-kin individuals than those hosts parasitized by siblings. Kin discrimination probably occurs by self-referent phenotype matching, where an individual compares its own phenotype with that of a non-familiar related individual, a mechanism rarely demonstrated in animals. The label used to recognize kin from non-kin corresponds to substances contained in the plug placed on the hosts by the resident larvae during the parasitization process. Kin competition induced by a limited larval dispersion may have favoured the evolution of kin recognition in this solitary species.

Gut microbiota and kin recognition

The animal gut contains a large reservoir of symbionts. Whilst these microbes have obvious physiological functions in, for example, digestion and immune defence, they can also affect their hosts behavior. Increasing evidence suggests that gut microbiota alters the scent of an individual, thereby affecting mate choice and kin recognition.

Experience of mating rivals causes males to modulate sperm transfer in the fly Drosophila pseudoobscura.

Male responses to risk of sperm competition play an important role in sexual selection, sexual conflict, and the evolution of mating systems. Such responses can combine behavioural and physiological processes, and can be mediated through different components of the ejaculate such as sperm numbers and seminal proteins. An additional level of ejaculate complexity is sperm heteromorphism, with the inclusion of non-fertilising parasperm in the ejaculate. We now test the response to rivals in a sperm heteromorphic species, Drosophila pseudoobscura, measuring the behavioural response and sperm transfer and, crucially, relating these to short-term fitness. Males respond to exposure to conspecific rivals by increasing mating duration, but do not respond to heterospecific rivals. In addition, after exposure to a conspecific rival, males increased the transfer of fertilising eusperm, but not non-fertilising parasperm. Males exposed to a conspecific rival also achieve higher offspring production. This suggests that the evolution of parasperm in flies was not driven by sperm competition and adds to the increasing evidence that males can make extremely sophisticated responses to mating competition.

Perception of male–male competition influences Drosophila copulation behaviour even in species where females rarely remate

Males in many taxa are known to exhibit behavioural plasticity in response to the perceived intensity of sperm competition, reflected in Drosophila melanogaster by increased copulation duration following prior exposure to a rival. We tested the prediction that males do not adjust their copulation effort in response to the presence of a competitor in Drosophila species where there is little or no sperm competition. Contrary to expectations, male plasticity in copulation duration was found in both Drosophila subobscura and Drosophila acanthoptera, species in which females rarely remate. These results are discussed in relation to the adaptive basis of plasticity in these species.

Extreme cost of rivalry in a monandrous species: male-male interactions result in failure to acquire mates and reduced longevity.

Mating system variation is profound in animals. In insects, female willingness to remate varies from mating with hundreds of males (extreme polyandry) to never remating (monandry). This variation in female behaviour is predicted to affect the pattern of selection on males, with intense pre-copulatory sexual selection under monandry compared to a mix of pre- and post-copulatory forces affecting fitness under polyandry. We tested the hypothesis that differences in female mating biology would be reflected in different costs of pre-copulatory competition between males. We observed that exposure to rival males early in life was highly costly for males of a monandrous species, but had lower costs in the polyandrous species. Males from the monandrous species housed with competitors showed reduced ability to obtain a mate and decreased longevity. These effects were specific to exposure to rivals compared with other types of social interactions (heterospecific male and mated female) and were either absent or weaker in males of the polyandrous species. We conclude that males in monandrous species suffer severe physiological costs from interactions with rivals and note the significance of male–male interactions as a source of stress in laboratory culture.

Kin Recognition in Aleochara bilineata Could Support the Kinship Theory of Genomic Imprinting

Genomic imprinting corresponds to the differential expression of a gene according to its paternal or maternal origin. The kinship theory of genomic imprinting proposes that maternally or paternally inherited genes may be in conflict over their effects on kin differently related along the paternal or maternal line. Most examples supporting the kinship theory of imprinting deal with competition between offspring for maternal resources. However, genomic imprinting may also explain differential behavioral expression toward kin whenever sibs are more related to each other via one parental sex than the other. Unfortunately, nothing is currently known about imprinting associated with a behavioral phenotype in insects. Here we report the first evidence of such a maternally imprinted behavior. We show that the solitary parasitoid larvae of Aleochara bilineata Gyll (Coleoptera; Staphylinidae), which avoid superparasitizing their full sibs, also avoid their cousins when they are related to them through their father, but not when they are related to them through their mother. A genetic kin recognition mechanism is proposed to explain this result and we conclude that genomic imprinting could control the avoidance of kin superparasitism in this species and have a profound influence on decision-making processes.

Males do not prolong copulation in response to competitor males in the polyandrous fly Drosophila bifasciata

Males in many taxa exhibit behavioural plasticity in response to the perceived threat of sperm competition. Drosophila males prolong mating in response to the presence of competitor males before copulation. The benefits of this behaviour to males are evident in Drosophila melanogaster. However, the adaptive nature of the trait is challenged by the observation that it is present in four other Drosophila species, two of which are largely monandrous, raising the possibility that this plasticity is not evolutionarily labile. In the present study, behavioural plasticity and the mating system in Drosophila bifasciata Pominini (Diptera, Drosophilidae) are analyzed. By contrast to other Drosophila species, male D. bifasciata do not exhibit plasticity in copulation duration when competitor males are present before mating. Thus, plasticity in mating duration is not fixed in the genus Drosophila. The mating system of D. bifasciata is also examined. The species is polyandrous but, uncommonly for the genus Drosophila, males transfer a mating plug composed of sperm to females, which forms very shortly after copulation and fills the female uterus. The absence of plasticity observed in D. bifasciata may arise from the sperm plug.

Assessment of Rival Males through the Use of Multiple Sensory Cues in the Fruitfly Drosophila pseudoobscura

Environments vary stochastically, and animals need to behave in ways that best fit the conditions in which they find themselves. The social environment is particularly variable, and responding appropriately to it can be vital for an animal’s success. However, cues of social environment are not always reliable, and animals may need to balance accuracy against the risk of failing to respond if local conditions or interfering signals prevent them detecting a cue. Recent work has shown that many male Drosophila fruit flies respond to the presence of rival males, and that these responses increase their success in acquiring mates and fathering offspring. In Drosophila melanogaster males detect rivals using auditory, tactile and olfactory cues. However, males fail to respond to rivals if any two of these senses are not functioning: a single cue is not enough to produce a response. Here we examined cue use in the detection of rival males in a distantly related Drosophila species, D. pseudoobscura, where auditory, olfactory, tactile and visual cues were manipulated to assess the importance of each sensory cue singly and in combination. In contrast to D. melanogaster, male D. pseudoobscura require intact olfactory and tactile cues to respond to rivals. Visual cues were not important for detecting rival D. pseudoobscura, while results on auditory cues appeared puzzling. This difference in cue use in two species in the same genus suggests that cue use is evolutionarily labile, and may evolve in response to ecological or life history differences between species.

Kin recognition loss following anesthesia in beetle larvae (Aleochara bilineata, Coleoptera, Staphylinidae)

Kin recognition is a complex cognitive process, where an individual should detect a phenotypic cue and compare it to an internal template, which might be genetically determined (i.e., innate or acquired). Kin recognition mechanisms will depend on whether previous encounters with kin are possible or not to form the individual internal template. When relatives have never met before, kin recognition is supposed to rely on recognition alleles (which allows the innate recognition of relatives bearing them), or on self-referent phenotype matching (the individual has formed a template using its own phenotype and recognizes as kin individuals which match it closely enough). Although self-referent phenotype matching is in theory the more likely explanation, it has not been possible so far to exclude experimentally the recognition alleles’ hypothesis. Here, we report that kin recognition in the solitary parasitoid larvae of Aleochara bilineata (Coleoptera; Staphylinidae) is suppressed following carbon dioxide anesthesia or chill-coma, both treatments known to cause a temporary amnesia. Treated larvae superparasitize indifferently hosts parasitized either by siblings or by non-kin larvae, while untreated larvae avoid hosts occupied by siblings. The two types of anesthesia thus suppress kin recognition, but their global effect on larvae is different. Chill-coma suppresses the ability to distinguish parasitized from unparasitized hosts and reduces parasitism rate, suggesting an aspecific impairment of sensory receptors or cognition. However, carbon dioxide narcosis only impairs kin recognition, strongly suggesting that an intact memory is necessary for kin recognition to take place. Although this study does not address the recognition alleles’ hypothesis per se, our results strongly support a self-referent phenotype matching mechanism. On the whole, kin recognition in A. bilineata larvae is effective through short-term memory, because it is affected by amnesic treatments.