Integrative characterization of genetic and phenotypic differentiation in an ant species complex with strong hierarchical population structure and low dispersal abilities.

Low dispersal, occurrence of asexual reproduction and geographic discontinuity increase genetic differentiation between populations, which ultimately can lead to speciation. In this work, we used a multidisciplinary framework to characterize the genetic and phenotypic differentiation between and within two cryptic ant species with restricted dispersal, Cataglyphis cursor and C. piliscapa and used behavioral experiments to test for reproductive isolation. Their distribution is segregated by the Rhône River and they have been traditionally distinguished only by hair numbers, although a statistical assessment is still lacking. We found strong genetic (microsatellites, nuclear and mitochondrial sequences), morphological (number of hairs, tibia length, male genitalia) and chemical (cuticular hydrocarbons) differentiation not only between species but also among localities within species. However, inter-specific differentiation was slightly higher than intra-specific differentiation for most markers. Overall, this pattern could either reflect reproductive isolation or could result from a longer period of geographic isolation between species than among localities within species without necessarily involving reproductive isolation. Interestingly, our behavioral experiments showed an absence of mating between species associated to a higher aggressiveness of workers towards heterospecific males. This suggests that sexual selection may, at least partially, fuel reproductive isolation. We also showed that cuticular hydrocarbons, mtDNA sequences and number of hairs provide reliable criteria allowing species discrimination. Overall, this species complex offers a case study to further investigate varying stages of a speciation continuum by estimating reproductive isolation between pairs of localities varying by their level of genetic differentiation.

Spring colonies of the ant Temnothorax nylanderi tolerate cadmium better than winter colonies, in both a city and a forest habitat.

A recent study showed that, in the ant Temnothorax nylanderi, city colonies are more tolerant to cadmium than forest colonies. However, because of annual variation in biological factors (e.g. body size, anti-stress protein production or trace metal accumulation rate), trace metal tolerance may vary over the year. We aimed at testing whether tolerance to cadmium of colonies of T. nylanderi differs between two different seasons within the same year (winter and spring). We also assessed whether the better cadmium tolerance of city colonies was constant over these two different time points. We collected colonies at the end of their hibernation period (winter colonies) and several weeks after (spring colonies) from two different habitats (forest and city) to assess whether response to cadmium was consistent regardless of the environment. We exposed colonies to a cadmium or a control treatment for 61 days. We compared tolerance to cadmium between spring/winter and city/forest colonies by measuring several life history traits. We found that spring colonies tolerates cadmium better than winter colonies, and that city colonies have a higher tolerance to cadmium but only in spring. Although further studies with replicated pairs of city/forest habitats and different years will be necessary to confirm those results, our study suggests that tolerance to trace metals can fluctuate along the yearly cycle.

Urbanization without isolation: the absence of genetic structure among cities and forests in the tiny acorn ant Temnothorax nylanderi.

Urban alteration of neutral and adaptive evolutionary processes is still underexplored. Using a genome-wide SNP dataset, we investigated (i) urban-induced modifications of population demography, genetic diversity and population structure and (ii) signature of divergent selection between urban and forest populations in the ant species, Temnothorax nylanderi. Our results did not reveal an impact of urbanization on neutral processes since we observed: (i) analogous genetic diversity among paired urban/forest sites and two control populations; (ii) weak population genetic structure explained neither by habitat (urban versus forest) nor by geography; (iii) a remarkably similar demographic history across populations with an ancestral growth followed by a recent decline, regardless of their current habitat or geographical location. The micro-geographical home range of ants may explain their resilience to urbanization. Finally, we detected 19 candidate loci discriminating urban/forest populations and associated with core cellular components, molecular function or biological process. Two of these loci were associated with a gene ontology term that was previously found to belong to a module of co-expressed genes related to caste phenotype. These results call for transcriptomics analyses to identify genes associated with ant social traits and to infer their potential role in urban adaptation.

Facultative use of thelytokous parthenogenesis for queen production in the polyandrous ant Cataglyphis cursor.

The evolutionary paradox of sex remains one of the major debates in evolutionary biology. The study of species capable of both sexual and asexual reproduction can elucidate factors important in the evolution of sex. One such species is the ant Cataglyphis cursor, where the queen maximizes the transmission of her genes by producing new queens (gynes) asexually while simultaneously maintaining a genetically diverse workforce via the sexual production of workers. We show that the queen can also produce gynes sexually and may do so to offset the costs of asexual reproduction. We genotyped 235 gynes from 18 colonies and found that half were sexually produced. A few colonies contained both sexually and asexually produced gynes. Although workers in this species can also use thelytoky, we found no evidence of worker production of gynes based on genotypes of 471 workers from the six colonies producing sexual gynes. Gynes are thus mainly, and potentially exclusively, produced by the queen. Simulations of gynes inbreeding level following one to ten generations of automictic thelytoky suggest that the queen switches between or combines thelytoky and sex, which may reduce the costs of inbreeding. This is supported by the relatively small size of inbred gynes in one colony, although we found no relationship between the level of inbreeding and immune parameters. Such facultative use of sex and thelytoky by individual queens contrasts with other known forms of parthenogenesis in ants, which are typically characterized by distinct lineages specializing in one strategy or the other.

Phenotypic plasticity of immune defence linked with foraging activity in the ant Cataglyphis velox.

Because immune defences are costly, life-history theories predict a modulation of immune investment according to its potential benefits. Social insects provide interesting models since infection risk may vary among individuals within a colony. In particular, the foraging workers, that have to leave the nest, suffer a higher infection risk and can contaminate their nest, which may favour high immune investments. However, evolutionary theories of aging predict that foragers should reduce their immune investment when they suffer high extrinsic mortality. To test these two predictions, we investigated the levels of phenoloxidase (PO) and prophenoloxidase, two important enzymes of the insect immune system, in workers of the ant Cataglyphis velox. We found a higher PO activity in foragers than in intra-nidal workers. This could result from an adaptive upregulation of the harmful PO (an enzyme potentially leading to autoimmune reactions) only when the risk of infection and wounding is high.

Hierarchical analysis of population genetic structure in the monogynous ant Cataglyphis cursor using microsatellite and mitochondrial DNA markers.

Despite having winged queens, female dispersal in the monogynous ant Cataglyphis cursor is likely to be restricted because colonies reproduce by fission. We investigated the pattern of population genetic structure of this species using eight microsatellite markers and a mitochondrial DNA (mtDNA) sequence, in order to examine the extent of female and nuclear gene flow in two types of habitat. Sampling was carried out at a large spatial scale (16 sites from 2.5 to 120 km apart) as well as at a fine spatial scale (two 4.5-km transects, one in each habitat type). The strong spatial clustering of mtDNA observed at the fine spatial scale strongly supported a restricted effective female dispersal. In agreement, patterns of the mtDNA haplotypes observed at large and fine spatial scales suggested that new sites are colonized by nearby sites. Isolation by distance and significant nuclear genetic structure have been detected at all the spatial scales investigated. The level of local genetic differentiation for mitochondrial marker was 15 times higher than for the nuclear markers, suggesting differences in dispersal pattern between the two sexes. However, male gene flow was not sufficient to prevent significant nuclear genetic differentiation even at short distances (500 m). Isolation-by-distance patterns differed between the two habitat types, with a linear decrease of genetic similarities with distance observed only in the more continuous of the two habitats. Finally, despite these low dispersal capacities and the potential use of parthenogenesis to produce new queens, no signs of reduction of nuclear genetic diversity was detected in C. cursor populations.

Very low genetic variability in the Indian queenless ant Diacamma indicum.

Abstract We developed microsatellite markers and combined them with mitochondrial markers to analyse the population genetic structure of the queenless ant Diacamma indicum. This species, lacking winged queens, is likely to have a restricted female dispersal but exhibits various life history traits suggesting higher dispersal abilities than the other Diacamma species. Only 4 of 11 microsatellites were polymorphic and only 1 had more than 4 alleles over 166 individuals originating from 7 populations from the south of India. Only one mitochondrial DNA (mtDNA) haplotype was detected throughout India (including one population in the north) and Sri Lanka. Such a level of polymorphism is particularly low compared with other Diacamma species having much smaller ranges in the south of India. A strong genetic differentiation was observed between populations separated by more than a few kilometres. We also analysed the genetic differentiation between the Indian populations and two populations from the Japanese island of Okinawa, which are morphologically similar and might belong to the same species. The genetic differentiation was high for both markers, suggesting an absence of ongoing gene flow between these populations.

The degree of extra-pair paternity increases with genetic variability.

The amount of extra-pair paternity in socially monogamous bird species varies from 0% to 76% extra-pair offspring. The causes of this remarkable interspecific variation are largely unknown, although intraspecific analyses suggest that females seek extra-pair matings to improve the genetic quality of their offspring. If this is a general explanation for the occurrence of extra-pair matings, then proportionally more females should seek to modify the paternity of their clutch when there is more variation among males in their genetic quality. Here we test this prediction in birds and show that interspecific variation in the proportion of extra-pair offspring is positively related to the proportion of polymorphic loci as measured by protein electrophoresis, even when controlling for potentially confounding variables. Genetic variability was also assessed, for sister pairs of species and populations differing significantly in extra-pair paternity, by using random priming, which provides an estimate of genome-wide diversity. We found that genetic diversity was higher in the populations with a higher level of extra-pair paternity. These results suggest that the amount of genetic variability in a population may be an important factor influencing mating patterns.

The genetical and environmental determination of phally polymorphism in the freshwater snail Bulinus truncatus.

In some species of self-fertile pulmonate snails, two sexual morphs co-occur in natural populations: regular individuals and aphallic individuals that cannot transmit sperm to other snails. Purely aphallic populations therefore reproduce obligatorily by selfing. Understanding the evolution of aphally and selfing in these snails requires a precise knowledge of phally determination. In this paper, we investigate the genetic and environmental determination of aphally in Bulinus truncatus by a survey of the family (offspring) aphally ratio of 233 individuals originating from seven natural populations and a study of the reaction norm of the family aphally ratio to temperature using 60 individuals from 10 selfed lineages of one population. Our results indicate a high genetic variability for the determination of aphally between populations and within some populations, associated with a high level of genetic determination. Our second experiment indicates a significant temperature and lineage effect though no interaction between these two effects. We discuss our results in the framework of threshold models developed for dimorphic traits with polygenic inheritance. We propose that the sexual morph of an individual at a given temperature is determined by a temperature threshold value depending on both the individual genotype and probabilistic processes.