Rapid molecular evolution of Spiroplasma symbionts of Drosophila

Spiroplasma are a group of Mollicutes whose members include plant pathogens, insect pathogens, and endosymbionts of animals. Spiroplasma phenotypes have been repeatedly observed to be spontaneously lost in Drosophila cultures, and several studies have documented a high genomic turnover in Spiroplasma symbionts and plant pathogens. These observations suggest that Spiroplasma evolves quickly in comparison to other insect symbionts. Here, we systematically assess evolutionary rates and patterns of Spiroplasma poulsonii, a natural symbiont of Drosophila. We analysed genomic evolution of sHy within flies, and sMel within in vitro culture over several years. We observed that S. poulsonii substitution rates are among the highest reported for any bacteria, and around two orders of magnitude higher compared with other inherited arthropod endosymbionts. The absence of mismatch repair loci mutS and mutL is conserved across Spiroplasma and likely contributes to elevated substitution rates. Further, the closely related strains sMel and sHy (>99.5% sequence identity in shared loci) show extensive structural genomic differences, which potentially indicates a higher degree of host adaptation in sHy, a protective symbiont of Drosophila hydei. Finally, comparison across diverse Spiroplasma lineages confirms previous reports of dynamic evolution of toxins, and identifies loci similar to the male-killing toxin Spaid in several Spiroplasma lineages and other endosymbionts. Overall, our results highlight the peculiar nature of Spiroplasma genome evolution, which may explain unusual features of its evolutionary ecology.

Thermal sensitivity of the Spiroplasma-Drosophila hydei protective symbiosis: The best of climes, the worst of climes

The outcome of natural enemy attack in insects has commonly been found to be influenced by the presence of protective symbionts in the host. The degree to which protection functions in natural populations, however, will depend on the robustness of the phenotype to variation in the abiotic environment. We studied the impact of a key environmental parameter – temperature – on the efficacy of the protective effect of the symbiont Spiroplasma on its host Drosophila hydei, against attack by the parasitoid wasp Leptopilina heterotoma. In addition, we investigated the thermal sensitivity of the symbiont’s vertical transmission, which may be a key determinant of the ability of the symbiont to persist. We found that vertical transmission was more robust than previously considered, with Spiroplasma being maintained at 25 °C, 18 °C and with 18/15 °C diurnal cycles, with rates of segregational loss only increasing at 15 °C. Protection against wasp attack was ablated before symbiont transmission was lost, with the symbiont failing to rescue the fly host at 18 °C. We conclude that the presence of a protective symbiosis in natural populations cannot be simply inferred from presence of a symbiont whose protective capacity has been tested under narrow controlled conditions. More broadly, we argue that the thermal environment is likely to represent an important determinant of the evolutionary ecology of defensive symbioses in natural environments, potentially driving seasonal, latitudinal and altitudinal variation in symbiont frequency, and modulating the strength of selection for symbiotic protective systems compared to defensive systems encoded in the nuclear genomes.

Mite choice generates sex- and size-biased infection in Drosophila hydei

SUMMARYHeterogeneities in parasite infection among conspecific hosts often manifest as sex- or size-biased infections, which are typically attributed to differential host susceptibility and exposure. Since parasite fitness is often tied to host quality, host preference by parasites is likely to be under strong selection. We test the hypothesis that host preference is sufficient to generate variability in infection rate among conspecifics. Specifically, we ask whether the mite Macrocheles muscaedomesticae is able to discriminate between Drosophila hydei hosts of different sex and size, while explicitly accounting for the potential confounding effects of these two factors. Our results indicate a preference for female hosts, but this preference appears to be driven by size and not sex per se. When differences in body size were controlled for, the sex-biased infection disappeared, while mites presented with the choice of two female flies of disparate sizes were more likely to select the larger host. Across the distribution of fly body weight in this study, mites preferentially attached to flies of intermediate size. This study provides evidence that mite choice for certain host types can play an important role in parasite transmission, even in the absence of differential susceptibility or exposure among hosts.

Sex-specific differences in the physiological basis of water conservation in the fruit fly Drosophila hydei from the western Himalayas

In the cosmopolitan fruit fly Drosophila hydei Sturtevant, 1921 (Diptera: Drosophilidae), the relative abundance of males is significantly higher than females, but the physiological basis of such sex-specific differences are largely unknown. For wild populations of D. hydei, we found seasonal changes (summer versus autumn) in desiccation-related traits, but the desiccation tolerance of males was higher than that of females in all seasons. For desiccation-related traits, we tested whether thermal developmental acclimation at three temperatures (17, 21, and 28 °C) matched seasonal changes observed under wild conditions. Male flies showed significantly higher trait values for desiccation resistance, cuticular lipid mass, hemolymph content, carbohydrate content, and dehydration tolerance compared with females when reared at lower or higher temperatures despite the lack of significant sex-specific differences in the total body-water content of flies reared at a particular growth temperature. We observed plastic changes in the amount of cuticular lipids consistent with corresponding differences in the rate of water loss. Treatment of cuticular surface with organic solvent (hexane) supported the role of cuticular lipids in affecting transcuticular water loss. We found significant thermal plastic effects for desiccation-related traits of D. hydei, but the sexual dimorphism was in the opposite direction, i.e., males were more desiccation resistant than females in D. hydei, whereas the reverse is true for many other Drosophila species. Our results suggest that sex-specific differences in the level of desiccation resistance in D. hydei are good predictors of relative abundance levels of male and female flies under wild conditions.