Testing for genetic assimilation with phylogenetic comparative analysis: Conceptual, methodological, and statistical considerations

Genetic assimilation is a process that leads to reduced phenotypic plasticity during adaptation to novel conditions, a potentially important phenomenon under global environmental change. Null expectations when testing for genetic assimilation, however, are not always clear. For instance, the statistical artifact of regression to the mean could bias us towards detecting genetic assimilation when it has not occurred. Likewise, the specific mechanism underlying plasticity expression may affect null expectations under neutral evolution. We used macroevolutionary numerical simulations to examine both of these important issues and their interaction, varying whether or not plasticity evolves, the evolutionary mechanism, trait measurement error, and experimental design. We also modified an existing reaction norm correction method to account for phylogenetic non-independence. We found: 1) regression to the mean is pervasive and can generate spurious support for genetic assimilation; 2) experimental design and post-hoc correction can minimize this spurious effect; and 3) neutral evolution can produce patterns consistent with genetic assimilation without constraint or selection, depending on the mechanism of plasticity expression. Additionally, we re-analyzed published macroevolutionary data supporting genetic assimilation, and found that support was lost after proper correction. Considerable caution is thus required whenever investigating genetic assimilation and reaction norm evolution at macroevolutionary scales.

Turtle Nest-Site Choice, Anthropogenic Challenges, and Evolutionary Potential for Adaptation

Oviparous animals, such as turtles, lay eggs whose success or demise depends on environmental conditions that influence offspring phenotype (morphology, physiology, and in many reptiles, also sex determination), growth, and survival, while in the nest and post-hatching. Consequently, because turtles display little parental care, maternal provisioning of the eggs and female nesting behavior are under strong selection. But the consequences of when and where nests are laid are affected by anthropogenic habitat disturbances that alter suitable nesting areas, expose eggs to contaminants in the wild, and modify the thermal and hydric environment experienced by developing embryos, thus impacting hatchling survival and the sexual fate of taxa with temperature-dependent sex determination (TSD) and genotypic sex determination (GSD). Indeed, global and local environmental change influences air, water, and soil temperature and moisture, which impact basking behavior, egg development, and conditions within the nest, potentially rendering current nesting strategies maladaptive as offspring mortality increases and TSD sex ratios become drastically skewed. Endocrine disruptors can sex reverse TSD and GSD embryos alike. Adapting to these challenges depends on genetic variation, and little to no heritability has been detected for nest-site behavior. However, modest heritability in threshold temperature (above and below which females or males develop in TSD taxa, respectively) exists in the wild, as well as interpopulation differences in the reaction norm of sex ratio to temperature, and potentially also in the expression of gene regulators of sexual development. If this variation reflects additive genetic components, some adaptation might be expected, provided that the pace of environmental change does not exceed the rate of evolution. Research remains urgently needed to fill current gaps in our understanding of the ecology and evolution of nest-site choice and its adaptive potential, integrating across multiple levels of organization.

Do Differences in Latitudinal Distributions of Species and Organelle Haplotypes Reflect Thermal Reaction Norms Within the Emiliania/Gephyrocapsa Complex?

The cosmopolitan phytoplankter Emiliania huxleyi contrasts with its closest relatives that are restricted to narrower latitudinal bands, making it interesting for exploring how alternative outcomes in phytoplankton range distributions arise. Mitochondrial and chloroplast haplogroups within E. huxleyi are shared with their closest relatives: Some E. huxleyi share organelle haplogroups with Gephyrocapsa parvula and G. ericsonii which inhabit lower latitudes, while other E. huxleyi share organelle haplogroups with G. muellerae, which inhabit high latitudes. We investigated whether the phylogeny of E. huxleyi organelles reflects environmental gradients, focusing on the Southeast Pacific where the different haplogroups and species co-occur. There was a high congruence between mitochondrial and chloroplast haplogroups within E. huxleyi. Haplogroup II of E. huxleyi is negatively associated with cooler less saline waters, compared to haplogroup I, both when analyzed globally and across temporal variability at the small special scale of a center of coastal upwelling at 30° S. A new mitochondrial haplogroup Ib detected in coastal Chile was associated with warmer waters. In an experiment focused on inter-species comparisons, laboratory-determined thermal reaction norms were consistent with latitudinal/thermal distributions of species, with G. oceanica exhibiting warm thermal optima and tolerance and G. muellerae exhibiting cooler thermal optima and tolerances. Emiliania huxleyi haplogroups I and II tended to exhibit a wider thermal niche compared to the other Gephyrocapsa, but no differences among haplogroups within E. huxleyi were found. A second experiment, controlling for local adaptation and time in culture, found a significant difference between E. huxleyi haplogroups. The difference between I and II was of the expected sign, but not the difference between I and Ib. The differences were small (≤1°C) compared to differences reported previously within E. huxleyi by local adaptation and even in-culture evolution. Haplogroup Ib showed a narrower thermal niche. The cosmopolitanism of E. huxleyi might result from both wide-spread generalist phenotypes and specialist phenotypes, as well as a capacity for local adaptation. Thermal reaction norm differences can well explain the species distributions but poorly explain distributions among mitochondrial haplogroups within E. huxleyi. Perhaps organelle haplogroup distributions reflect historical rather than selective processes.

Multiple trait breeding programs with genotype-by-environment interactions based on reaction norms, with application to genetic improvement of disease resilience

Background Genotype-by-environment interactions for a trait can be modeled using multiple-trait, i.e. character-state, models, that consider the phenotype as a different trait in each environment, or using reaction norm models based on a functional relationship, usually linear, between phenotype and a quantitative measure of the quality of the environment. The equivalence between character-state and reaction norm models has been demonstrated for a single trait. The objectives of this study were to extend the equivalence of the reaction norm and character-state models to a multiple-trait setting and to both genetic and environmental effects, and to illustrate the application of this equivalence to the design and optimization of breeding programs for disease resilience. Methods Equivalencies between reaction norm and character-state models for multiple-trait phenotypes were derived at the genetic and environmental levels, which demonstrates how multiple-trait reaction norm parameters can be derived from multiple-trait character state parameters. Methods were applied to optimize selection for a multiple-trait breeding goal in a target environment based on phenotypes collected in a healthy and disease-challenged environment, and to optimize the environment in which disease-challenge phenotypes should be collected. Results and conclusions The equivalence between multiple-trait reaction norm and multiple-trait character-state parameters allow genetic improvement for a multiple-trait breeding goal in a target environment to be optimized without recording phenotypes and estimating parameters for the target environment.

Mothers front-load their investment to the egg stage when helped in a wild cooperative bird

In many cooperative societies, including our own, helpers assist with the post-natal care of breeders' young, and may thereby benefit the post-natal development of offspring. Here we present evidence of a novel mechanism by which such post-natal helping could also have hitherto unexplored beneficial effects on pre-natal development: by lightening post-natal maternal workloads, helpers may allow mothers to increase their pre-natal investment per offspring. We present the findings of a decade-long study of cooperatively breeding white-browed sparrow weaver, Plocepasser mahali, societies. Within each social group, reproduction is monopolized by a dominant breeding pair, and non-breeding helpers assist with nestling feeding. Using a within-mother reaction norm approach to formally identify maternal plasticity, we demonstrate that when mothers have more female helpers they decrease their own post-natal investment per offspring (feed their nestlings at lower rates) but increase their pre-natal investment per offspring (lay larger eggs, which yield heavier hatchlings). That these plastic maternal responses are predicted by female helper number, and not male helper number, implicates the availability of post-natal helping per se as the likely driver (rather than correlated effects of group size), because female helpers feed nestlings at substantially higher rates than males. We term this novel maternal strategy 'maternal front-loading' and hypothesize that the expected availability of post-natal help allows helped mothers to focus maternal investment on the pre-natal phase, to which helpers cannot contribute directly. Such cryptic maternally mediated helper effects on pre-natal development may markedly complicate attempts to identify and quantify the fitness consequences of helping.

Genotype-environment interaction effects on weight gain in cattle using reaction norms

The existence of genotype-environment interaction (GEI) using reaction norm models and their impact on the genetic evaluation of Nellore sires for body weight at 120, 210, 365 and 450 days of age was verified. Three models were used: animal model (AM) that disregards GEI and the one-step reaction norm model with homogeneous and heterogeneous residual variance (1SRNMH_het). Bayes Inference via Gibbs Sampling was used to estimate the variance components. The AM model better fits to weights at 120 and 210 days of age, while 1SRNMH_het was more adequate for body weights at 365 and 450 days of age, suggesting the existence of GEI. The posterior means of direct heritability were 0.33±0.01 and 0.36±0.01 and maternal heritability of 0.21±0.01 and 0.19±0.01 for body weights at 120 and 210 days of age, respectively. For body weights at 365 and 450 days of age, posterior means of heritability varied along the environmental gradient, but the ranking of sires based on breeding values was not changed by different environmental gradients. All rank correlations were greater than 0.80, strongly suggesting a scale effect of GEI. Despite the evidence of GEI on post-weaning weight gain, it did not change the ranking of sires. Therefore, it did not have a relevant impact on the genetic evaluation of sires because they are robust to environmental changes.

PSVII-27 Genetic analysis of disease resilience of nursery-to-finish pigs under a natural disease challenge model using reaction norms

Disease resilience is the ability of an animal to maintain performance across environments with different disease challenge loads (CL) and can be quantified using random regression reaction norm models that describe phenotype as a function of CL. Objectives of this study were to: 1) develop measures of CL using growth rate and clinical disease phenotypes under a natural disease challenge; 2) evaluate genetic variation in disease resilience. Data used were late nursery and finisher growth rates and clinical disease phenotypes, including medical treatment and mortality rates, and subjective health scores, collected on 50 batches of 60/75 crossbred (LRxY) barrows under a polymicrobial natural disease challenge. All pigs were genotyped using a 650K SNP panel. Different CL were derived from estimates of contemporary group effects and used as environmental covariates in reaction norm analyses of average daily gain (ADG) and treatment rate (TRT). The CL were compared based on model loglikelihoods and estimates of genetic variance, using both linear and cubic spline reaction norm models. Linear reaction norm models fitted the data significantly better than the standard genetic model and the cubic spline models fitted the data significantly better than the linear reaction norm model for most traits. CL based on early finisher ADG provided the best fit for nursery ADG, while CL based on clinical disease phenotypes was best for finisher ADG and TRT. With increasing CL, estimates of heritability for ADG initially decreased and then increased, while estimates of heritability for TRT generally increased with CL. Genetic correlations were low between ADG or TRT at high versus low CL but high for close CLs. Results can be used to select more resilient pigs across different CL levels, or high-performance animals at a given CL level, or a combination of these. Funded by Genome Canada, Genome Alberta, USDA-NIFA, and PigGenCanada.

Urbanization extends flight phenology and leads to local adaptation of seasonal plasticity in Lepidoptera

Urbanization is gaining force globally, which challenges biodiversity, and it has recently also emerged as an agent of evolutionary change. Seasonal phenology and life cycle regulation are essential processes that urbanization is likely to alter through both the urban heat island effect (UHI) and artificial light at night (ALAN). However, how UHI and ALAN affect the evolution of seasonal adaptations has received little attention. Here, we test for the urban evolution of seasonal life-history plasticity, specifically changes in the photoperiodic induction of diapause in two lepidopterans, Pieris napi (Pieridae) and Chiasmia clathrata (Geometridae). We used long-term data from standardized monitoring and citizen science observation schemes to compare yearly phenological flight curves in six cities in Finland and Sweden to those of adjacent rural populations. This analysis showed for both species that flight seasons are longer and end later in most cities, suggesting a difference in the timing of diapause induction. Then, we used common garden experiments to test whether the evolution of the photoperiodic reaction norm for diapause could explain these phenological changes for a subset of these cities. These experiments demonstrated a genetic shift for both species in urban areas toward a lower daylength threshold for direct development, consistent with predictions based on the UHI but not ALAN. The correspondence of this genetic change to the results of our larger-scale observational analysis of in situ flight phenology indicates that it may be widespread. These findings suggest that seasonal life cycle regulation evolves in urban ectotherms and may contribute to ecoevolutionary dynamics in cities.