As organisms age, the effectiveness of natural selection weakens, leading to age-related decline in fitness-related traits. The evolution of age-related changes associated with senescence is likely influenced by mutation accumulation (MA) and antagonistic pleiotropy (AP). MA predicts that age-related decline in fitness components is driven by age-specific sets of alleles, nonnegative genetic correlations within trait across age, and an increase in the coefficient of genetic variance. AP predicts that age-related decline in a trait is driven by alleles with positive effects on fitness in young individuals and negative effects in old individuals, and is expected to lead to negative genetic correlations within traits across age. We build on these predictions using an association mapping approach to investigate the change in additive effects of SNPs across age and among traits for multiple stress-response fitness-related traits, including cold stress with and without acclimation and starvation resistance. We found support for both MA and AP theories of aging in the age-related decline in stress tolerance. Our study demonstrates that the evolution of age-related decline in stress tolerance is driven by a combination of alleles that have age-specific additive effects, consistent with MA, as well as nonindependent and antagonistic genetic architectures characteristic of AP.

Understanding how thermal selection affects phenotypic distributions across different time scales will allow us to predict the effect of climate change on the fitness of ectotherms. We tested how seasonal temperature variation affects basal levels of cold tolerance and two types of phenotypic plasticity in Drosophila melanogaster. Developmental acclimation occurs as developmental stages of an organism are exposed to seasonal changes in temperature and its effect is irreversible, while reversible short-term acclimation occurs daily in response to diurnal changes in temperature. We collected wild flies from a temperate population across seasons and measured two cold tolerance metrics (chill-coma recovery and cold stress survival) and their responses to developmental and short-term acclimation. Chill-coma recovery responded to seasonal shifts in temperature, and phenotypic plasticity following both short-term and developmental acclimation improved cold tolerance. This improvement indicated that both types of plasticity are adaptive, and that plasticity can compensate for genetic variation in basal cold tolerance during warmer parts of the season when flies tend to be less cold tolerant. We also observed a significantly stronger trade-off between basal cold tolerance and short-term acclimation during warmer months. For the longer-term developmental acclimation, a trade-off persisted regardless of season. A relationship between the two types of plasticity may provide additional insight into why some measures of thermal tolerance are more sensitive to seasonal variation than others. K E Y W O R D Sacclimation, daily temperature, developmental plasticity, evolutionary constraint, seasonal temperature

A range of heavy metals are required for normal cell function and homeostasis. However, the anthropogenic release of metal compounds into soil and water sources presents a pervasive health threat. Copper is one of many heavy metals that negatively impacts diverse organisms at a global scale. Using a combination of quantitative trait locus (QTL) mapping and RNA sequencing in the Drosophila Synthetic Population Resource, we demonstrate that resistance to the toxic effects of ingested copper in D. melanogaster is genetically complex and influenced by allelic and expression variation at multiple loci. QTL mapping identified several QTL that account for a substantial fraction of heritability. Additionally, we find that copper resistance is impacted by variation in behavioral avoidance of copper and may be subject to life-stage specific regulation. Gene expression analysis further demonstrated that resistant and sensitive strains are characterized by unique expression patterns. Several of the candidate genes identified via QTL mapping and RNAseq have known copper-specific functions (e.g., Ccs, Sod3, CG11825), and others are involved in the regulation of other heavy metals (e.g., Catsup, whd). We validated several of these candidate genes with RNAi suggesting they contribute to variation in adult copper resistance. Our study illuminates the interconnected roles that allelic and expression variation, organism life stage, and behavior play in copper resistance, allowing a deeper understanding of the diverse mechanisms through which metal pollution can negatively impact organisms.

We report on the discovery of a remarkable defensive specialization in stonefishes that was identified during a phylogenetic study of scorpionfishes and their relatives. This newly described innovation, the lachrymal saber, involves modifications to the circumorbitals, maxilla, adductor mandibulae, and associated tendons. At its core, the lachrymal saber is an elongation of an anterior spine (or spines) on the ventral surface of the lachrymal that stonefishes are capable of rotating from the standard ventral position to a locked lateral position. The locking mechanism minimally includes a bony spur on the inner surface of the lachrymal and a ridged bony protuberance on the anterolateral end of the maxilla. A modified and highly subdivided adductor mandibulae appears to control the movement of the lachrymal saber by rotating the maxilla while it is engaged with the spur on the medial side of the lachrymal. This maxillary rotation results in a subsequent rotation of the lachrymal that we hypothesize reduces predation on stonefishes. This specialization was included in our phylogenetic analysis of scorpaenoid fishes. This study expands upon the previous higher-level taxonomic sampling reported in earlier evolutionary studies of scorpaenoid fishes and, unlike previous analyses, explicitly combines molecular and morphological data with an expanded taxonomic sampling to mitigate the conflict between these competing datasets. The resulting phylogeny based on a combination of 113 morphological and 5,280 molecular characters for 63 species is used to produce a revised taxonomy of flatheads, scorpionfishes, sea robins, and stonefishes. Our results do not support the monophyly of the traditional Scorpaeniformes, Scorpaenoidei, Scorpaenoidea, Platycephaloidea, Bembridae, Scorpaenidae, Sebastidae, Serranidae, Tetrarogidae, or Triglidae. Our monophyletic taxonomy recognizes nine monophyletic families: Bembridae, Congiopodidae, Hoplichthyidae, Neosebastidae, Platycephalidae, Plectrogeniidae, Scorpaenidae, Synanceiidae, and Triglidae. The taxonomic composition of the Congiopodidae, Hoplichthyidae, Neosebastidae, and Platycephalidae are unchanged. The Bembridae is expanded to include the recently described Parabembridae, while Bembradium is moved to the Plectrogeniidae. The Scorpaenidae is expanded to include the traditional Sebastidae and Setarchidae. The Triglidae is expanded to include the Peristediidae. Finally, a revised Synanceiidae, diagnosed by the lachrymal saber, is expanded to include the Apistidae, Aploactinidae, Eschmeyeridae, Gnathanacanthidae, Pataecidae, Perryenidae, and Tetrarogidae. Based on these results, we recommend treating all of these traditional scorpaenoid clades as families in an expanded Scorpaeniformes that includes a restricted Scorpaenoidei that includes all traditional scorpaenoid families except the Congiopodidae. The resulting phylogeny is then used to explore aspects of scorpaenoid evolution.

The ability to survive periods without food is an important component of individual fitness, and genetic dissection can provide insight into the mechanisms and evolution of starvation resistance. Everman et al. use several genome-wide...

Drosophila community composition is complex in temperate regions with different abundance of flies and species across the growing season. Monitoring Drosophila populations provides insights into the phenology of both native and invasive species. Over a single growing season, we collected Drosophila at regular intervals and determined the number of individuals of the nine species we found in Kansas, USA. Species varied in their presence and abundance through the growing season with peak diversity occurring after the highest seasonal temperatures. We developed models for the abundance of the most common species, Drosophila melanogaster , D . simulans , D . algonquin , and the recent invasive species, D . suzukii . These models revealed that temperature played the largest role in abundance of each species across the season. For the two most commonly studied species, D . melanogaster and D . simulans , the best models indicate shifted thermal optima compared to laboratory studies, implying that fluctuating temperature may play a greater role in the physiology and ecology of these insects than indicated by laboratory studies, and should be considered in global climate change studies.

Environmental challenges presented by temperature variation can be overcome through phenotypic plasticity in small invasive ectotherms. We tested the effect of thermal exposure to 21, 18, and 11°C throughout the whole life cycle of individuals, thermal exposure of adults reared at 25°C to 15 and 11°C for a 21-d period, and long (14:10 hr) and short (10:14 hr) photoperiod on ovary size and development in Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) cultured from a recently established population in Topeka, Kansas (United States). Examination of the response to temperature and photoperiod variation in this central plains population provides insight into the role of phenotypic plasticity in a climate that is warmer than regions in North America where D. suzukii was initially established. We found both low temperature and short photoperiod resulted in reduced ovary size and level of development. In particular, reduced ovary development was observed following exposure to 15°C, indicating that ovary development in females from the central plains population is more sensitive to lower temperature compared with populations examined from the northern United States and southern Canada. We also provide evidence that D. suzukii reared at 25°C are capable of short-term hardening when exposed to -6°C following 4°C acclimation, contrary to previous reports indicating flies reared at warm temperatures do not rapidly-cold harden. Our study highlights the central role of phenotypic plasticity in response to winter-like laboratory conditions and provides an important geographic comparison to previously published assessments of ovary development and short-term hardening survival response for D. suzukii collected in cooler climates.

Fitness is determined by the ability of an organism to both survive and reproduce; however, the mechanisms that lead to increased survival may not have the same effect on reproductive success. We used nineteen natural Drosophila melanogaster genotypes from the Drosophila Genetic Reference Panel to determine if adaptive plasticity following short-term acclimation through rapid cold-hardening (RCH) affects mating behavior and mating success. We confirmed that exposure to the acclimation temperature is beneficial to survival following cold stress; however, we found that this same acclimation temperature exposure led to less efficient male courtship and a significant decrease in the likelihood of mating. Cold tolerance and the capacity to respond plastically to cold stress were not correlated with mating behavior following acclimation, suggesting that the genetic control of the physiological effects of the cold temperature exposure likely differ between survival and behavioral responses. We also tested whether the exposure of males to the acclimation temperature influenced courtship song. This exposure again significantly increased courtship duration; however, courtship song was unchanged. These results illustrate costs of short-term acclimation on survival and reproductive components of fitness and demonstrate the pronounced effect that short-term thermal environment shifts can have on reproductive success.