Body size trends in response to climate and urbanization in the widespread North American deer mouse, Peromyscus maniculatus

© 2020, The Author(s). Body size decline is hypothesized to be a key response to climate warming, including warming driven by urban heat islands. However, urbanization may also generate selective gradients for body size increases in smaller endotherms via habitat fragmentation. Here we utilize a densely sampled, multi-source dataset to examine how climate and urbanization affect body size of Peromyscus maniculatus (PEMA), an abundant rodent found across North America. We predicted PEMA would conform to Bergmann’s Rule, e.g. larger individuals in colder climates, spatially and temporally. Hypotheses regarding body size in relation to urbanization are less clear; however, with increased food resources due to greater anthropogenic activity, we expected an increase in PEMA size. Spatial mixed-models showed that PEMA conform to Bergmann’s Rule and that PEMA were shorter in more urbanized areas. With the inclusion of decade in mixed-models, we found PEMA mass, but not length, is decreasing over time irrespective of climate or population density. We also unexpectedly found that, over time, smaller-bodied populations of PEMA are getting larger, while larger-bodied populations are getting smaller. Our work highlights the importance of using dense spatiotemporal datasets, and modeling frameworks that account for bias, to better disentangle broad-scale climatic and urbanization effects on body size

Hybridization between the Woodland Salamanders Plethodon cinereus and P. electromorphus Is Not Widespread

A recent study reported widespread hybridization between the Eastern Red-backed Salamander (Plethodon cinereus) and the Northern Ravine Salamander (P. electromorphus) in northern Ohio. In this study, DNA sequence data were obtained from three nuclear loci and 20 single nucleotide polymorphisms (SNPs) were identified from the sequences. They found that 48 out of 90 individuals from 13 populations were hybrids, and in some localities every individual possessed an admixed genotype. As these results contradict our observations, and because levels of hybridization impact our interpretation of past and ongoing studies, we revisited the data. First we reanalyzed the original SNPs using STRUCTURE, then we repeated the analysis using haplotypes instead of SNPs. We found that K1⁄4 2 was best supported by both analyses, and they agree in recovering lower levels of hybridization than originally reported. For example, five populations in the original study identified as highly admixed or composed entirely of admixed genotypes we found to be pure P. cinereus or to lack evidence of extensive admixture. Similar results were obtained using NEWHYBRIDS and analyses based on gene trees. We conclude that while hybridization between P. cinereus and P. electromorphus occurs, it is much more restricted than originally reported

Genetic Analysis of a Cryptic Contact Zone between Mitochondrial Clades of the Eastern Red-Backed Salamander, Plethodon cinereus

When evolutionarily divergent lineages adjoin their geographic ranges after a period of isolation, myriad outcomes can occur, from population anastomosis to the evolution of reproductive isolation by way of reinforcement. Hybrid zones represent natural experiments that may indicate whether lineages will maintain their evolutionary independence. Here, we report on a hybrid zone in the Eastern Red-Backed Salamander, Plethodon cinereus, a highly abundant and wide-ranging terrestrial salamander found in the northeastern United States and in southeastern Canada. An earlier study identified six distinct mitochondrial clades across the range of P. cinereus. Populations of two of these clades were as close as 9.6 km apart in Lorain County, Ohio, USA. To investigate the nature of this contact zone, we sampled 316 individuals from 16 sites along a 53-km transect, and analyzed 10 microsatellite loci and one mitochondrial locus. We found a clinal transition for mtDNA haplotypes. In contrast, most studies of terrestrial plethodontid salamanders commonly exhibit sharp boundaries between mtDNA clades. Microsatellite markers, however, revealed little differentiation and weak population structure, suggesting the nuclear cline, if it exists, lies outside of our sampling region. Explanations for the discordance between the mitochondrial DNA and our microsatellite data include lineage sorting, male-biased dispersal, or historical introgression of mtDNA, among other possibilities. We compare our results with other studies of introgression in terrestrial salamanders, and discuss the causes of mitonuclear discordance

Do genetic structure and landscape heterogeneity impact color morph frequency in a polymorphic salamander?

Landscape heterogeneity plays an important role in population structure and divergence, particularly for species with limited vagility. Here, we used a landscape genetic approach to identify how landscape and environmental variables affect genetic structure and color morph frequency in a polymorphic salamander. The eastern red- backed salamander, Plethodon cinereus, is widely distributed in northeastern North America and contains two common color morphs, striped and unstriped, that are divergent in ecology, behavior, and physiology. To quantify population structure, rates of gene flow, and genetic drift, we amplified 10 microsatellite loci from 648 individuals across 28 sampling localities. This study was conducted in northern Ohio, where populations of P. cinereus exhibit an unusually wide range of morph frequency variation. To test whether genetic distance was more correlated with morph frequency, elevation, canopy cover, waterways, ecological niche or geographic distance, we used resistance distance and least cost path analyses. We then examined whether landscape and environmental variables, genetic distance or geographic distance were correlated with variation in morph frequency. Tests for population structure revealed three genetic clusters across our sampling range, with one cluster monomorphic for the striped morph. Rates of gene flow and genetic drift were low to moderate across sites. Genetic distance was most correlated with ecological niche, elevation and a combination of landscape and environmental variables. In contrast, morph frequency variation was correlated with waterways and geographic distance. Thus, our results suggest that selection is also an important evolutionary force across our sites, and a balance between gene flow, genetic drift and selection interact to maintain the two color morphs

Colour scales with climate in North American ratsnakes: a test of the thermal melanism hypothesis using community science images

Animal colour is a complex trait shaped by multiple selection pressures that can vary across geography. The thermal melanism hypothesis predicts that darker coloration is beneficial to animals in colder regions because it allows for more rapid solar absorption. Here, we use community science images of three closely related species of North American ratsnakes (genus Pantherophis) to examine if climate predicts colour variation across range-wide scales. We predicted that darker individuals are found in colder regions and higher elevations, in accordance with the thermal melanism hypothesis. Using an unprecedented dataset of over 8000 images, we found strong support for temperature as a key predictor of darker colour, supporting thermal melanism. We also found that elevation and precipitation are predictive of colour, but the direction and magnitude of these effects were more variable across species. Our study is the first to quantify colour variation in Pantherophis ratsnakes, highlighting the value of community science images for studying range-wide colour variation

Dietary alkaloid sequestration in a poison frog:\ud an experimental test of alkaloid uptake in Melanophryniscus\ud stelzneri (Bufonidae)

Several lineages of brightly colored anurans independently evolved the ability to secrete alkaloid-containing defensive chemicals from granular glands in the skin. These species, collectively referred to as ‘poison frogs,’ form a polyphyletic assemblage that includes some species of Dendrobatidae, Mantellidae, Myobatrachidae, Bufonidae, and Eleutherodactylidae. The ability to sequester alkaloids from dietary arthropods has been demonstrated experimentally in most poison frog lineages but not in bufonid or eleutherodactylid poison frogs. As with other poison frogs, species of the genus Melanophryniscus (Bufonidae) consume large numbers of mites and ants, suggesting they might also sequester defensive alkaloids from dietary sources. To test this hypothesis, fruit flies dusted with alkaloid/nutritional supplement powder were fed to individual Melanophryniscus stelzneri in two experiments. In the first experiment, the alkaloids 5,8-disubstituted indolizidine 235B' and decahydroquinoline were administered to three individuals for 104 days. In the second experiment, the alkaloids 3,5-disubstituted indolizidine 239Q and decahydroquinoline were given to three frogs for 153 days. Control frogs were fed fruit flies dusted only with nutritional supplement. Gas chromatography/mass spectrometry analyses revealed that skin secretions of all experimental frogs contained alkaloids, whereas those of all control frogs lacked alkaloids. Uptake of decahydroquinoline was greater than uptake of 5,8-disubstituted indolizidine, and uptake of 3,5-disubstituted indolizidine was greater than uptake of decahydroquinoline, suggesting greater uptake efficiency of certain alkaloids. Frogs in the second experiment accumulated a greater amount of alkaloid, which corresponds to the longer duration and greater number of alkaloid-dusted fruit flies that were consumed. These findings provide the first experimental evidence that bufonid poison frogs sequester alkaloid-based defenses from dietary sourcesWe thank V. Korb for her assistance in frog feeding, and M.A. Nichols for invaluable assistance in maintaining the GC/MS instrument. We extend a special thanks to C.D. Anthony, C. Hickerson, and C.A. Sheil for providing suggestions and comments that improved the quality of this manuscript.TG was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico Proc. 307001/ 2011-3 and Fundação de Amparo à Pesquisa do Estado de São Paulo Proc. 2012/10000-5

Methods for broad-scale plant phenology assessments using citizen scientists’ photographs

© 2020 Barve et al. Applications in Plant Sciences is published by Wiley Periodicals, Inc. on behalf of the Botanical Society of America Premise: Citizen science platforms for sharing photographed digital vouchers, such as iNaturalist, are a promising source of phenology data, but methods and best practices for use have not been developed. Here we introduce methods using Yucca flowering phenology as a case study, because drivers of Yucca phenology are not well understood despite the need to synchronize flowering with obligate pollinators. There is also evidence of recent anomalous winter flowering events, but with unknown spatiotemporal extents. Methods: We collaboratively developed a rigorous, consensus-based approach for annotating and sharing whole plant and flower presence data from iNaturalist and applied it to Yucca records. We compared spatiotemporal flowering coverage from our annotations with other broad-scale monitoring networks (e.g., the National Phenology Network) in order to determine the unique value of photograph-based citizen science resources. Results: Annotations from iNaturalist were uniquely able to delineate extents of unusual flowering events in Yucca. These events, which occurred in two different regions of the Desert Southwest, did not appear to disrupt the typical-period flowering. Discussion: Our work demonstrates that best practice approaches to scoring iNaturalist records provide fine-scale delimitation of phenological events. This approach can be applied to other plant groups to better understand how phenology responds to changing climate

Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium

The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-m porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal versus disease-related endometrial microenvironments