Molecular examination of representatives of Balaustium from several populations in SW Poland, performed using the sequence data from the mitochondrial cytochrome c oxidase subunit I, confirmed their common specific affiliation and identity with Balaustium murorum. The potential presence of distinct species in the studied material, preliminarily inferred from the discovery of clusters as a result of Principal Component Analysis exploring the metric data sets, was rejected due to the finding of only one haplotype, at intra- and inter-population sampling. An insight into meristic traits in larvae, focused on chaetotaxy of legs, revealed wider variation than hitherto recognized for the species. The variation was higher in laboratory-reared larvae compared to field-collected ones. The overall deviations from the mean character values at intra- and interpopulation levels, higher than hitherto observed for the species, vote for the reappraisal of the criteria adopted for discrimination of members of Balaustium with the application of an integrative approach.

Living organisms adapt to their environment, and this adaptive response to environmental changes is influenced by both genomic and epigenomic components. As adaptation underpins tolerance to stressors, it is crucial to consider biological adaptation in evaluating the adverse outcomes of environmental chemicals, such as biocides. Daphnid studies have revealed differences in sensitivity to environmental chemicals between conspecific populations or clones, as well as between species. This study aimed to identify whether sensitivity to chemicals is subject to intraspecific variation, and whether this sensitivity depends on the genetic and epigenetic backgrounds of the daphnid population. We used an integrative approach to assess the comparative toxicity of a mixture of 5-chloro-2-methyl-4-isothiazoline-3-one and 2-methyl-4-isothiazolin-3-one (CMIT/MIT), a commonly used isothiazolinone biocide, by measuring mortality, reproduction, physiological traits, global DNA methylation, and proteomic expression at the species and strain levels. The results showed that the variation in sensitivity to CMIT/MIT between conspecific strains (Daphnia pulex; DPR vs. DPA strains) could exceed that observed between congeneric species (D. magna vs. D. pulex DPR strain). Under the control conditions, DPR (the strain most sensitive to CMIT/MIT) was characterized by a larger body size, a higher heart rate, and a higher level of global DNA methylation compared to its counterpart (DPA), and proteome profiles differed between the two strains. Particularly, the study identified strain-specific epigenetic and proteomic responses to LC20 of CMIT/MIT, demonstrating putative critical proteins and biological pathways associated with the observed differences in phenotype and sensitivity to CMIT/MIT. Downregulation of certain proteins (e.g., SAM synthase, GSTs, hemoglobin, and cuticle proteins) and DNA hypomethylation can be proposed as key events (KEs) of adverse outcome pathway (AOP) for isothiazolinone toxicity. Our findings indicate that both genetic variations and epigenetic modifications can lead to intraspecific variation in sensitivity to chemicals, and this variation should be considered in the ecological risk assessment framework for chemical substances. We suggest conducting further analysis on methylated gene regions and observing transgenerational effects to verify the role of crosstalk between genetic and epigenetic factors in phenotypic and protein expressions. DATA AVAILABILITY: Proteomic data is available in supplementary materials.

The reasons why some species occur widespread, while related species have restricted geographical ranges have been attributed to habitat specialization or ecological niche breadth. For species in the genus Saxifraga, habitat specialization alone cannot explain the distributional differences observed. We hypothesize that recruitment traits (i.e., germination, emergence, and survival) may account for differences in geographical ranges and that early life stages correlate to survival. We studied recruitment responses in 13 widespread and 12 narrow-ranged Saxifraga species in the laboratory and common garden experiments using seeds collected from 79 populations in the European Alps. We found that in the laboratory cold temperature led to higher germination percentages compared with warm temperature for both distribution groups. This represents an exception to the general assumption that alpine species require warm cues for germination. In warm laboratory temperatures, widespread species germinated better than narrow-ranged species, indicating a greater tolerance of warm temperatures for the former. Subsequent to germination, recruitment traits between the two distribution groups were lower or null in the common garden, suggesting that the impact of recruitment on species\' geographical ranges occurs at the earliest life stage. Mean time to emergence of narrow-ranged species showed lower variability than that of widespread species. Consistently, intraspecific variation of mean annual temperatures between seed collection sites was lower for narrow-ranged species, indicating a close relationship between home sites and emergence time. Emergence percentage was a strong predictor of survival only for widespread species, underlining that seed and seedling functional traits differ between distribution groups, which require further research. Our results support the view that early life stages are critical to population dynamics and thus can influence species\' geographical ranges. The wider responses to climatic conditions in widespread species may have facilitated their spread across the Alps. Our results also suggest that all Saxifraga species face a considerable threat from climate warming due to their overall cold-adapted recruitment niche.

Generally, the species is considered to be the only naturally occurring taxon. However, species recognised and defined using different species delimitation criteria cannot readily be compared, impacting studies of biodiversity through Deep Time. This comparability issue is particularly marked when comparing extant with extinct species, because the only available data for species delimitation in fossils is derived from their preserved morphology, which is generally restricted to osteology in vertebrates. Here, we quantify intraspecific, intrageneric, and intergeneric osteological variability in extant species of lacertid lizards using pairwise dissimilarity scores based on a dataset of 253 discrete osteological characters for 99 specimens referred to 24 species. Variability is always significantly lower intraspecifically than between individuals belonging to distinct species of a single genus, which is in turn significantly lower than intergeneric variability. Average values of intraspecific variability and associated standard deviations are consistent (with few exceptions), with an overall average within a species of 0.208 changes per character scored. Application of the same methods to six extinct lacertid species (represented by 40 fossil specimens) revealed that intraspecific osteological variability is inconsistent, which can at least in part be attributed to different researchers having unequal expectations of the skeletal dissimilarity within species units. Such a divergent interpretation of intraspecific and interspecific variability among extant and extinct species reinforces the incomparability of the species unit. Lacertidae is an example where extant species recognised and defined based on a number of delimitation criteria show comparable and consistent intraspecific osteological variability. Here, as well as in equivalent cases, application of those skeletal dissimilarity values to palaeontological species delimitation potentially provides a way to ameliorate inconsistencies created by the use of morphology to define species.

Organism shape changes predictably during ontogeny, resulting in specific patterns of ontogenetic allometry. In several plant and animal lineages, among-species variation in the shape of mature organisms mirrors variation along their growth trajectories. Hence, ontogenetic allometry is an important bias in evolution. This bias should be stronger at reduced evolutionary time scales, in which among-trait correlations had less time to evolve. Nevertheless, it was shown that adaptation of organism shape frequently involved departures from the ancestral ontogenetic allometry. Moreover, only a moderate fraction of shape variation is correlated with size during ontogeny. Hence, nonallometric variation in shape (NAVSh) is likely to contribute to adaptation, even at reduced evolutionary time scales. We explored the contributions of allometric variation in shape (AVSh), NAVSh, and size variation to adaptive evolution in the angiosperm species Calceolaria polyrhiza. This strongly relies on oil-collecting bees for pollination and experienced transitions in the size of pollinators during the last 2 Ma. Using geometric morphometrics, we described corolla morphology in several populations across its distribution range. Variation in corolla shape was decomposed into an allometric and a nonallometric component, and corolla size was estimated. We then looked for the correlation between these aspects of morphology and the pollinator. Our results suggest that adaptation to pollinators with different sizes relied on NAVSh, which resulted from shifts in the allometric slope and from shape changes that occurred early in flower development. We conclude that NAVSh can contribute to adaptation in flowering plants, even at the species-level.

Body temperature is a key factor in physiological processes, influencing lizard performances; and life history traits are expected to generate variability of thermal preferences in different individuals. Gender, body size and reproductive condition may impose specific requirements on preferred body temperatures. If these three factors have different physiological functions and thermal requirements, then the preferred temperature may represent a compromise that optimizes these physiological functions. Therefore, the body temperatures that lizards select in a controlled environment may reflect a temperature that maximizes their physiological needs. The tegu lizard Tupinambis merianae is one of the largest lizards in South America and has wide ontogenetic variation in body size and sexual dimorphism. In the present study we evaluate intraspecific variability of thermal preferences of T. merianae. We determined the selected body temperature and the rate at which males and females attain their selected temperature, in relation to body size and reproductive condition. We also compared the behavior in the thermal gradient between males and females and between reproductive condition of individuals. Our study show that T. merianae selected body temperature within a narrow range of temperatures variation in the laboratory thermal gradient, with 36.24±1.49°C being the preferred temperature. We observed no significant differences between sex, body size and reproductive condition in thermal preferences. Accordingly, we suggest that the evaluated categories of T. merianae have similar thermal requirements. Males showed higher rates to obtain heat than females and reproductive females, higher rates than non-reproductive ones females. Moreover, males and reproductive females showed a more dynamic behavior in the thermal gradient. Therefore, even though they achieve the same selected temperature, they do it differentially.

Intraspecific variability in social systems is gaining increased recognition in primatology. Many primate species display variability in pair-living social organizations through incorporating extra adults into the group. While numerous models exist to explain primate pair-living, our tools to assess how and why variation in this trait occurs are currently limited. Here I outline an approach which: (i) utilizes conceptual models to identify the selective forces driving pair-living; (ii) outlines novel possible causes for variability in social organization; and (iii) conducts a holistic species-level analysis of social behavior to determine the factors contributing to variation in pair-living. A case study on white-faced sakis (Pithecia pithecia) is used to exemplify this approach. This species lives in either male-female pairs or groups incorporating \"extra\" adult males and/or females. Various conceptual models of pair-living suggest that high same-sex aggression toward extra-group individuals is a key component of the white-faced saki social system. Variable pair-living in white-faced sakis likely represents alternative strategies to achieve competency in this competition, in which animals experience conflicting selection pressures between achieving successful group defense and maintaining sole reproductive access to mates. Additionally, independent decisions by individuals may generate social variation by preventing other animals from adopting a social organization that maximizes fitness. White-faced saki inter-individual relationships and demographic patterns also lend conciliatory support to this conclusion. By utilizing both model-level and species-level approaches, with a consideration for potential sources of variation, researchers can gain insight into the factors generating variation in pair-living social organizations.