Inferring the history of speciation in house mice from autosomal,X‐linked,Y‐linked and mitochondrial genes

Patterns of genetic differentiation among taxa at early stages of divergence provide an opportunity to make inferences about the history of speciation. Here, we conduct a survey of DNA‐sequence polymorphism and divergence at loci on the autosomes, X chromosome, Y chromosome and mitochondrial DNA in samples of Mus domesticus, M. musculus and M. castaneus. We analyzed our data under a divergence with gene flow model and estimate that the effective population size of M. castaneus is 200 000–400 000, of M. domesticus is 100 000–200 000 and of M. musculus is 60 000–120 000. These data also suggest that these species started to diverge approximately 500 000 years ago. Consistent with this recent divergence, we observed considerable variation in the genealogical patterns among loci. For some loci, all alleles within each species formed a monophyletic group, while at other loci, species were intermingled on the phylogeny of alleles. This intermingling probably reflects both incomplete lineage sorting and gene flow after divergence. Likelihood ratio tests rejected a strict allopatric model with no gene flow in comparisons between each pair of species. Gene flow was asymmetric: no gene flow was detected into M. domesticus, while significant gene flow was detected into both M. castaneus and M. musculus. Finally, most of the gene flow occurred at autosomal loci, resulting in a significantly higher ratio of fixed differences to polymorphisms at the X and Y chromosomes relative to autosomes in some comparisons, or just the X chromosome in others, emphasizing the important role of the sex chromosomes in general and the X chromosome in particular in speciation.     

Tracing carbon and oxygen isotope signals from newly assimilated sugars in the leaves to the tree-ring archive

The analysis of δ ¹³C and δ ¹⁸O in tree-ring archives offers retrospective insights into environmental conditions and ecophysiological processes. While photosynthetic carbon isotope discrimination and evaporative oxygen isotope enrichment are well understood, we lack information on how the isotope signal is altered by downstream metabolic processes.
In Pinus sylvestris , we traced the isotopic signals from their origin in the leaf water ( δ ¹⁸O) or the newly assimilated carbon ( δ ¹³C), via phloem sugars to the tree-ring, over a time-scale that ranges from hours to a growing season.
Seasonally, variable ¹³C enrichment of sugars related to phloem loading and transport did lead to uncoupling between δ ¹³C in the tree-ring, and the c _i/ c _a ratio at the leaf level. In contrast, the oxygen isotope signal was transferred from the leaf water to the tree-ring with an expected enrichment of 27‰, with time-lags of approximately 2 weeks and with a 40% exchange between organic oxygen and xylem water oxygen during cellulose synthesis.
This integrated overview of the fate of carbon and oxygen isotope signals within the model tree species P. sylvestris provides a novel physiological basis for the interpretation of δ ¹³C and δ ¹⁸O in tree-ring ecology.     

DOES COLCHICINE AFFECT AGGREGATION OF NORMAL AND TRANSFORMED BHK CELLS DIFFERENTLY?

The effect of colchicine and vinblastine on cell aggregation was studied, using BHK cells and their transformed derivatives (pyBHK cells). When cells were dissociated with EDTA and the assay was made in a Ca²⁺-containing medium, the aggregation of transformed cells was prevented by colchicine and vinblastine, whereas the aggregation of normal cells was unaffected. When a Ca²⁺-free medium was used for aggregation, neither type of cell was influenced by these drugs. BHK and pyBHK cells, dissociated by trypsin in the presence of Ca²⁺, can aggregate only in the Ca²⁺-containing medium and the aggregation of both cell types was equally prevented by colchicine and vinblastine. Based on these results, it was concluded that colchicine and vinblastine inhibited the Ca²⁺-dependent mechanism of cell adhesion, but not the Ca²⁺-independent one which occurs in the Ca²⁺-free aggregation medium.     

Lack of photosynthetic or stomatal regulation after 9 years of elevated [CO2] and 4 years of soil warming in two conifer species at the alpine treeline

Alpine treelines are temperature‐limited vegetation boundaries. Understanding the effects of elevated [CO₂] and warming on CO₂ and H₂O gas exchange may help predict responses of treelines to global change. We measured needle gas exchange of Larix decidua Mill. and Pinus mugo ssp. uncinata DC trees after 9 years of free air CO₂ enrichment (575 µmol mol^?1) and 4 years of soil warming (+4 °C) and analysed δ¹³C and δ¹⁸O values of needles and tree rings. Tree needles under elevated [CO₂] showed neither nitrogen limitation nor end‐product inhibition, and no down‐regulation of maximal photosynthetic rate (A_max) was found. Both tree species showed increased net photosynthetic rates (A_n) under elevated [CO₂] (L. decidua: +39%; P. mugo: +35%). Stomatal conductance (g_H2O) was insensitive to changes in [CO₂], thus transpiration rates remained unchanged and intrinsic water‐use efficiency (iWUE) increased due to higher A_n. Soil warming affected neither A_n nor g_H2O. Unresponsiveness of g_H2O to [CO₂] and warming was confirmed by δ¹⁸O needle and tree ring values. Consequently, under sufficient water supply, elevated [CO₂] induced sustained enhancement in A_n and lead to increased C inputs into this ecosystem, while soil warming hardly affected gas exchange of L. decidua and P. mugo at the alpine treeline.     

Effects and Mitigation of Road Impacts on Individual Movement Behavior of Wildcats

ABSTRACT Roads can affect the persistence of wildlife populations, through posing mortality risks and acting as barriers. In many countries, transportation agencies attempt to counterbalance these negative impacts. Road mortality is a major threat for European wildcats (Felis silvestris); therefore, we tested the effectiveness of a newly developed wildcat-specific fence in preventing wildcat mortality along a new motorway. We hypothesized that such a fenced motorway would at the same time be a significant barrier to wildcats and may at worst result in 2 isolated populations. We used radiotracking data of 12 wildcats, resulting in 13,000 fixes, to investigate individual movement behavior during and after construction of a new motorway in southwestern Germany. The motorway was fenced with the wildcat-specific fence and included crossing structures, not especially constructed for wildlife. Additionally we collected road kills on stretches of the same motorway with various types of fencing. A rate of 0.4 wildcat kills/km/year on the motorway, which was traveled by 10,000 vehicles/day and fenced with a regular wildlife fence, was reduced by 83% on stretches with wildcat-specific fencing. Of the available crossing structures, wildcats preferred open-span viaducts. Road underpasses were used but hold a mortality risk themselves. As opposed to our expectations, the fenced motorway (fenced with wildcat fence) posed only a moderate barrier to wildcats. Individuals were hindered in their daily routine and some stopped crossing completely but others continued crossing regularly. The adaptation of spatial and temporal behavior to traffic volume and location of crossing structures has an energetic cost. Hence, we suggest that only a small number of major roads can be tolerated within a wildcat's home range. To meet the demands of the European Habitats Directive, we recommend installing the wildcat fence in wildcat core areas along motorways to reduce wildcat mortality. We suggest that fences should incorporate safe crossing structures every 1.5-2.5 km. Our findings in terms of fencing design and crossing structures can be used by transportation agencies for an effective reduction of road mortality and barrier effect for carnivores.     

How far do experimentally elevated CO2 levels reach into the surrounding? – An example using the 13C label of soil organic matter as an archive

During the last two decades, free air CO₂ enrichment (FACE) studies have been conducted to study the effects of rising atmospheric CO₂ concentrations on ecosystems. The distances between fumigated and control plots differ widely among those projects, but no experimental data are available how far into the surrounding area an effect of CO₂ fumigation can be detected. As the CO₂ gas added to the fumigated plots has a different ¹³C label than ambient atmospheric CO₂, its carbon can be traced into plants and soil organic matter (SOM). The Swiss FACE in Eschikon had been conducted for 10 years on a grassland site. After it had ended, we analysed soil samples from three transects extending from the plots to the surrounding area for their organic carbon (C_org) content and carbon isotopic signature. We determined the maximum spatial extension to which carbon originating from the fumigation was incorporated into SOM. A budget of the fumigation gas‐derived C_org in the upper 10 cm of the soil showed that approximately 50 kg C were stored within the plots, and an additional 31 kg C were stored in their immediate surroundings up to a distance of 9 m from the gas pipes. The presented approach provides us with a method to determine a posteriori the extension to which the CO₂ fumigation treatment contaminated its immediate surroundings during a FACE experiment. In the presented example, this showed that the distances between plots could have been reduced significantly. Although not generalizable to other experimental settings, the finding indicates that optimizing the spatial layout, e.g. by modelling gas dispersion, will be useful when planning future large‐scale FACE infrastructures. Our approach provides a solid basis to test such gas‐dispersion models on existing FACE sites before planning new sites.     

Development of microsatellite markers in the toxic dinoflagellate Alexandrium minutum (Dinophyceae)

Outbreaks of paralytic shellfish poisoning caused by the toxic dinoflagellate Alexandrium minutum (Dinophyceae) are a worldwide concern from both the economic and human health points of view. For population genetic studies of A. minutum distribution and dispersal, highly polymorphic genetic markers are of great value. We isolated 12 polymorphic microsatellites from this cosmopolitan, toxic dinoflagellate species. These loci provide one class of highly variable genetic markers, as the number of alleles ranged from four to 12, and the estimate of gene diversity was from 0.560 to 0.862 across the 12 microsatellites; these loci have the potential to reveal genetic structure and gene flow among A. minutum populations.     

Isolation of Zn2+ as an Endogenous Agonist of GPR39 from Fetal Bovine Serum

We attempted to determine natural agonists of GPR39 in fetal bovine serum (FBS). FBS was conditioned to extract peptides and fractionated by two types of HPLC. The activity of each fraction was monitored by intracellular calcium mobilization. Then the purified active ingredient was analyzed by inductively coupled plasma mass spectrometry. In this fashion, Zn^{2 +} ion was identified as an agonist of GPR39, though no peptidergic molecules were found. The calcium-mobilizing activity of Zn^{2 +} was not abolished by pertussis toxin but was by a phospholipase C (PLC) inhibitor, U73122, indicating that the activity of GPR39 is mediated through the G_qα -PLC pathway. In addition, Zn^{2 +} also activated mouse and rat GPR39, showing that the function of GPR39 as a Zn^{2 +} receptor is conserved across species. This study is the first exploration of GPR39 agonists in FBS and indicates that GPR39 functions as a Gq-coupled Zn^{2 +}-sensing receptor.