Behavioural and physiological adaptations to low-temperature environments in the common frog, Rana temporaria

<b>Background</b><p></p> Extreme environments can impose strong ecological and evolutionary pressures at a local level. Ectotherms are particularly sensitive to low-temperature environments, which can result in a reduced activity period, slowed physiological processes and increased exposure to sub-zero temperatures. The aim of this study was to assess the behavioural and physiological responses that facilitate survival in low-temperature environments. In particular, we asked: 1) do high-altitude common frog (Rana temporaria) adults extend the time available for larval growth by breeding at lower temperatures than low-altitude individuals?; and 2) do tadpoles sampled from high-altitude sites differ physiologically from those from low-altitude sites, in terms of routine metabolic rate (RMR) and freeze tolerance? Breeding date was assessed as the first day of spawn observation and local temperature recorded for five, paired high- and low-altitude R. temporaria breeding sites in Scotland. Spawn was collected and tadpoles raised in a common laboratory environment, where RMR was measured as oxygen consumed using a closed respiratory tube system. Freeze tolerance was measured as survival following slow cooling to the point when all container water had frozen.<p></p> <b>Results</b><p></p> We found that breeding did not occur below 5°C at any site and there was no significant relationship between breeding temperature and altitude, leading to a delay in spawning of five days for every 100 m increase in altitude. The relationship between altitude and RMR varied by mountain but was lower for individuals sampled from high- than low-altitude sites within the three mountains with the highest high-altitude sites (≥900 m). In contrast, individuals sampled from low-altitudes survived freezing significantly better than those from high-altitudes, across all mountains.<p></p> <b>Conclusions</b><p></p> Our results suggest that adults at high-altitude do not show behavioural adaptations in terms of breeding at lower temperatures. However, tadpoles appear to have the potential to adapt physiologically to surviving at high-altitude via reduced RMR but without an increase in freeze tolerance. Therefore, survival at high-altitude may be facilitated by physiological mechanisms that permit faster growth rates, allowing completion of larval development within a shorter time period, alleviating the need for adaptations that extend the time available for larval growth

Evolution of drug-tolerant nematode populations in response to density reduction

Resistance to xenobiotics remains a pressing issue in parasite treatment and global agriculture. Multiple factors may affect the evolution of resistance, including interactions between life-history traits and the strength of selection imposed by different drug doses. We experimentally created replicate selection lines of free-living Caenorhabditis remanei exposed to Ivermectin at high and low doses to assess whether survivorship of lines selected in drug-treated environments increased, and if this varied with dose. Additionally, we maintained lines where mortality was imposed randomly to control for differences in density between drug treatments and to distinguish between the evolutionary consequences of drug-treatment versus ecological processes due to changes in density-dependent feedback. After 10 generations, we exposed all of the selected lines to high-dose, low-dose and drug-free environments to evaluate evolutionary changes in survivorship as well as any costs to adaptation. Both adult and juvenile survival were measured to explore relationships between life-history stage, selection regime and survival. Intriguingly, both drug-selected and random-mortality lines showed an increase in survivorship when challenged with Ivermectin; the magnitude of this increase varied with the intensity of selection and life-history stage. Our results suggest that interactions between density-dependent processes and life history may mediate evolved changes in susceptibility to control measures

Combining molecular and incomplete observational data to inform management of southern white rhinoceros (Ceratotherium simum simum)

Conservation efforts have preserved the southern white rhinoceros (SWR) in protected areas and have resulted in substantial overall growth in population size, but in small, fragmented populations in which inbreeding is an important risk. However, field observation of breeding often lacks sufficient accuracy to inform translocation strategies that are intended to increase genetic variation. The purpose of this study was to integrate microsatellite genotypes with an incomplete, field-observed pedigree to make inferences about mean kinship and basic demographic data that could be used to inform translocation programmes for SWR in a confined population in Botswana. Using this approach, we identified parents for 29 out of 45 offspring born in the reserve between 1993 and 2013 and detected eight non-breeding bulls with high mean kinship as candidates for translocation. The method also allowed inferences about demographic parameters that could influence the effectiveness of intervention strategies, such as age and timing of reproduction, and natal sex ratios. Importantly, the reproductive dominance of the bulls was not as skewed as expected after the original dominant bull was removed from the population, suggesting that closed populations can maintain multiple, simultaneously breeding males. The genetic data also confirmed that the accuracy of field-based parentage assignment was increased after implementation of an ear-notching programme. This study demonstrates the value of combining genetic information with ongoing surveillance to inform management of threatened populations, and of using mean kinship to inform metapopulation management by identifying candidates for translocation

Changing environments and genetic variation: natural variation in inbreeding does not compromise short-term physiological responses

Selfing plant lineages are surprisingly widespread and successful in a broad range of environments, despite showing reduced genetic diversity, which is predicted to reduce their long-term evolutionary potential. However, appropriate short-term plastic responses to new environmental conditions might not require high levels of standing genetic variation. In this study, we tested whether mating system variation among populations, and associated changes in genetic variability, affected short-term responses to environmental challenges. We compared relative fitness and metabolome profiles of naturally outbreeding (genetically diverse) and inbreeding (genetically depauperate) populations of a perennial plant, Arabidopsis lyrata, under constant growth chamber conditions and an outdoor common garden environment outside its native range. We found no effect of inbreeding on survival, flowering phenology or short-term physiological responses. Specifically, naturally occurring inbreeding had no significant effects on the plasticity of metabolome profiles, using either multivariate approaches or analysis of variation in individual metabolites, with inbreeding populations showing similar physiological responses to outbreeding populations over time in both growing environments. We conclude that low genetic diversity in naturally inbred populations may not always compromise fitness or short-term physiological capacity to respond to environmental change, which could help to explain the global success of selfing mating strategies

Changing environments and genetic variation: natural variation in inbreeding does not compromise short-term physiological responses

Selfing plant lineages are surprisingly widespread and successful in a broad range of environments, despite showing reduced genetic diversity, which is predicted to reduce their long-term evolutionary potential. However, appropriate short-term plastic responses to new environmental conditions might not require high levels of standing genetic variation. In this study, we tested whether mating system variation among populations, and associated changes in genetic variability, affected short-term responses to environmental challenges. We compared relative fitness and metabolome profiles of naturally outbreeding (genetically diverse) and inbreeding (genetically depauperate) populations of a perennial plant, Arabidopsis lyrata, under constant growth chamber conditions and an outdoor common garden environment outside its native range. We found no effect of inbreeding on survival, flowering phenology or short-term physiological responses. Specifically, naturally occurring inbreeding had no significant effects on the plasticity of metabolome profiles, using either multivariate approaches or analysis of variation in individual metabolites, with inbreeding populations showing similar physiological responses to outbreeding populations over time in both growing environments. We conclude that low genetic diversity in naturally inbred populations may not always compromise fitness or short-term physiological capacity to respond to environmental change, which could help to explain the global success of selfing mating strategies

Sodalis glossinidius presence in wild tsetse is only associated with presence of trypanosomes in complex interactions with other tsetse-specific factors

Background: Susceptibility of tsetse flies (Glossina spp.) to trypanosomes of both humans and animals has been associated with the presence of the endosymbiont Sodalis glossinidius. However, intrinsic biological characteristics of the flies and environmental factors can influence the presence of both S. glossinidius and the parasites. It thus remains unclear whether it is the S. glossinidius or other attributes of the flies that explains the apparent association. The objective of this study was to test whether the presence of Trypanosoma vivax, T. congolense and T. brucei are related to the presence of S. glossinidius in tsetse flies when other factors are accounted for: geographic location, species of Glossina, sex or age of the host flies. Results: Flies (n = 1090) were trapped from four sites in the Shimba Hills and Nguruman regions in Kenya. Sex and species of tsetse (G. austeni, G. brevipalpis, G. longipennis and G. pallidipes) were determined based on external morphological characters and age was estimated by a wing fray score method. The presence of trypanosomes and S. glossinidius was detected using PCR targeting the internal transcribed spacer region 1 and the haemolysin gene, respectively. Sequencing was used to confirm species identification. Generalised Linear Models (GLMs) and Multiple Correspondence Analysis (MCA) were applied to investigate multivariable associations. The overall prevalence of trypanosomes was 42.1%, but GLMs revealed complex patterns of associations: the presence of S. glossinidius was associated with trypanosome presence but only in interactions with other factors and only in some species of trypanosomes. The strongest association was found for T. congolense, and no association was found for T. vivax. The MCA also suggested only a weak association between the presence of trypanosomes and S. glossinidius. Trypanosome-positive status showed strong associations with sex and age while S. glossinidius-positive status showed a strong association with geographic location and species of fly. Conclusions: We suggest that previous conclusions about the presence of endosymbionts increasing probability of trypanosome presence in tsetse flies may have been confounded by other factors, such as community composition of the tsetse flies and the specific trypanosomes found in different regions

Spatial variation in species composition of Saprolegnia, a parasitic oomycete of amphibian eggs, in Scotland

This document is the Accepted Manuscript version of a published work that appeared in final form in Herpetological Journal. To access the final edited and published work see http://www.thebhs.org/index.php?option=com_docman&Itemid=42.Parasitic water moulds in the genus Saprolegnia cause mortality of amphibian embryos and reduced size at metamorphosis, leading to increased adult mortality. Most studies of virulence have focused on only a single Saprolegnia species, but the Saprolegnia species associated with amphibian eggs and their distributions are not well known. This study aimed to investigate the distribution of amphibian-associated water moulds in Scotland. In particular, we asked the questions: i) Does Saprolegnia species composition vary between sites?; and ii) Is presence of Saprolegnia related to environmental parameters? Common frog (Rana temporaria) eggs with evidence of Saprolegnia infection were sampled from ten sites, cultured, and the 28S region of the rDNA array sequenced. Thirteen samples isolated from four sites were identified as members of the Saprolegniaceae and the ITS region of these samples were subsequently sequenced to further resolve species identification. Four species of Saprolegnia were found in total, with one or two species of Saprolegnia present in each of four sites. S. diclina was the most common species identified and was found at three of the four sites. Acidity was significantly lower and altitude significantly higher at sites where Saprolegniaceae were present. Therefore, R. temporaria eggs in different pools are subject to infection by different, and in some instances more than one, species of Saprolegnia. Overall, our findings suggest that future studies of virulence need to consider the effect of multiple Saprolegnia species within a site as well as the population of origin of the amphibian hostFieldwork was supported by grants from the Royal Geographic Society, the Glasgow Natural History Society and the Scottish Mountaineering Trust. This study was supported by PhD CASE studentship funding from the Biotechnology and Biological Sciences Research Council, in partnership with the Royal Zoological Society of Scotland

The maize INDETERMINATE1 flowering time regulator defines a highly conserved zinc finger protein family in higher plants

BACKGROUND: The maize INDETERMINATE1 gene, ID1, is a key regulator of the transition to flowering and the founding member of a transcription factor gene family that encodes a protein with a distinct arrangement of zinc finger motifs. The zinc fingers and surrounding sequence make up the signature ID domain (IDD), which appears to be found in all higher plant genomes. The presence of zinc finger domains and previous biochemical studies showing that ID1 binds to DNA suggests that members of this gene family are involved in transcriptional regulation. RESULTS: Comparison of IDD genes identified in Arabidopsis and rice genomes, and all IDD genes discovered in maize EST and genomic databases, suggest that ID1 is a unique member of this gene family. High levels of sequence similarity amongst all IDD genes from maize, rice and Arabidopsis suggest that they are derived from a common ancestor. Several unique features of ID1 suggest that it is a divergent member of the maize IDD family. Although no clear ID1 ortholog was identified in the Arabidopsis genome, highly similar genes that encode proteins with identity extending beyond the ID domain were isolated from rice and sorghum. Phylogenetic comparisons show that these putative orthologs, along with maize ID1, form a group separate from other IDD genes. In contrast to ID1 mRNA, which is detected exclusively in immature leaves, several maize IDD genes showed a broad range of expression in various tissues. Further, Western analysis with an antibody that cross-reacts with ID1 protein and potential orthologs from rice and sorghum shows that all three proteins are detected in immature leaves only. CONCLUSION: Comparative genomic analysis shows that the IDD zinc finger family is highly conserved among both monocots and dicots. The leaf-specific ID1 expression pattern distinguishes it from other maize IDD genes examined. A similar leaf-specific localization pattern was observed for the putative ID1 protein orthologs from rice and sorghum. These similarities between ID1 and closely related genes in other grasses point to possible similarities in function

RADseq and mate choice assays reveal unidirectional gene flow among three lamprey ecotypes despite weak assortative mating: Insights into the formation and stability of multiple ecotypes in sympatry

Adaptive divergence with gene flow often results in complex patterns of variation within taxa exhibiting substantial ecological differences among populations. One example where this may have occurred is the parallel evolution of freshwater‐resident nonparasitic lampreys from anadromous‐parasitic ancestors. Previous studies have focused on transitions between these two phenotypic extremes, but here, we considered more complex evolutionary scenarios where an intermediate freshwater form that remains parasitic is found sympatrically with the other two ecotypes. Using population genomic analysis (restriction‐associated DNA sequencing), we found that a freshwater‐parasitic ecotype was highly distinct from an anadromous‐parasitic form (Qlake‐P = 96.8%, Fst = 0.154), but that a freshwater‐nonparasitic form was almost completely admixed in Loch Lomond, Scotland. Demographic reconstructions indicated that both freshwater populations likely derived from a common freshwater ancestor. However, while the nonparasitic ecotype has experienced high levels of introgression from the anadromous‐parasitic ecotype (Qanad‐P = 37.7%), there is no evidence of introgression into the freshwater‐parasitic ecotype. Paradoxically, mate choice experiments predicted high potential for gene flow: Males from all ecotypes were stimulated to spawn with freshwater‐parasitic females, which released gametes in response to all ecotypes. Differentially fixed single nucleotide polymorphisms identified genes associated with growth and development, which could possibly influence the timing of metamorphosis, resulting in significant ecological differences between forms. This suggests that multiple lamprey ecotypes can persist in sympatry following shifts in adaptive peaks, due to environmental change during their repeated colonization of post‐glacial regions, followed by periods of extensive gene flow among such diverging populations

Local adaptation with high gene flow: temperature parameters drive adaptation to altitude in the common frog (Rana temporaria)

This is the peer reviewed version of the following article: Muir, A. P., Biek, R., Thomas, R. & Mable, B. K. (2014). Local adaptation with high gene flow: temperature parameters drive adaptation to altitude in the common frog (Rana temporaria). Molecular Ecology, 23(3), 561–574. DOI: 10.1111/mec.12624, which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1111/mec.12624/full. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-ArchivingBoth environmental and genetic influences can result in phenotypic variation. Quantifying the relative contributions of local adaptation and phenotypic plasticity to phenotypes is key to understanding the effect of environmental variation on populations. Identifying the selective pressures that drive divergence is an important, but often lacking, next step. High gene flow between high- and low-altitude common frog (Rana temporaria) breeding sites has previously been demonstrated in Scotland. The aim of this study was to assess whether local adaptation occurs in the face of high gene flow and to identify potential environmental selection pressures that drive adaptation. Phenotypic variation in larval traits was quantified in R. temporaria from paired high- and low-altitude sites using three common temperature treatments. Local adaptation was assessed using QST -FST analyses, and quantitative phenotypic divergence was related to environmental parameters using Mantel tests. Although evidence of local adaptation was found for all traits measured, only variation in larval period and growth rate was consistent with adaptation to altitude. Moreover, this was only evident in the three mountains with the highest high-altitude sites. This variation was correlated with mean summer and winter temperatures, suggesting that temperature parameters are potentially strong selective pressures maintaining local adaptation, despite high gene flow.Fieldwork was supported by grants from the Royal Geographic Society, the Glasgow Natural History Society and the Scottish Mountaineering Trust. Permission for sampling from protected areas was granted by Scottish Natural Heritage. This study was supported by PhD CASE studentship funding from the Biotechnology and Biological Sciences Research Council, in partnership with the Royal Zoological Society of Scotland. Reference