Cladogenesis of the European brown hare (Lepus europaeus Pallas, 1778)

A substantial portion of today’s biodiversity is attributed to the climatic oscillations of the Pleistocene Ice Ages. Gradual but dramatic climate changes were accompanied by expansion, contraction, and isolation of populations, promoting the accumulation of genome differences and adaptations in refugial populations and resulting in allopatric differentiation in a variety of taxa. In the present study, partial mitochondrial DNA sequences of the widely distributed European brown hare (Lepus europaeus) were analyzed to test whether the species’ present genetic structure is the result of postglacial re-colonization of Europe from Asia Minor (clade A) and the Balkans (clade B) only, as suggested previously, or if additional refugia are likely. Analyses indicated the presence of an additional refugium (Italy, clade I). The genealogic network of Italian hares displayed the tree-like structure expected from refugial populations, whereas central European brown hare haplotypes revealed a clear star-phylogeny indicative of past-bottleneck population growth. This population size expansion, which was confirmed by mismatch analysis, was estimated to have occurred ∼50–55 thousand years ago (kya). The divergence of clade A* from the remaining matrilines is estimated at 239 kya, whereas the divergence of the ancestors of clades B* and I from A* occurred about 128 kya. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Threatened but understudied: supporting conservation by understanding the genetic structure of the flat-headed cat

The flat-headed cat (Prionailurus planiceps) is a wetland specialist, currently facing habitat loss on a serious scale due to massive destruction of lowland forests and wetlands in Southeast Asia. Despite its ‘endangered’ status in the IUCN Red List, there has virtually been no investigation on the population structure nor on the evolutionary history of the flat-headed cat. To fill this gap, we used full mitochondrial genome sequences, obtained from archival samples, covering the historical distribution of the flat-headed cat. Our data revealed a high genetic differentiation (F _st = 0.81, P?<?0.001) between mitochondrial lineages from Borneo and those from Thai-Malay Peninsula/Sumatra, a split that was dated to ~575 Kya. Such a significant differentiation clearly distinguishes the Bornean flat-headed cat population from all other populations and suggests that there should be a reassessment of the flat-headed cat’s intraspecific taxonomy. However, morphological and nuclear data are required to corroborate our mtDNA results. Until such data become available, we recommend that for future conservation efforts and captive breeding programmes the two genetically distinct flat-headed cat populations are managed separately as two lineages to maintain the original genetic diversity of this endangered species.     

Evolution of caprine and ovine β-defensin genes

Defensins comprise an important family of anti-microbial peptides. Among vertebrates, numerous defensin genes have been detected, but their evolutionary background is still discussed. We investigated the molecular evolution and variability of β-defensins of Caprini via sequence analyses of defensin introns. Screening of several domestic and wild species of Caprini revealed a total of 13 discrete β-defensin coding sequences, with three of them described before this study. Phylogenetic analyses revealed that the array of newly described defensin genes is of monophyletic origin and has arisen in numerous independent duplication events after separation of the ancestral defensins. As a result of that scenario, recent defensin genes are distributed in a species-specific manner. Values of synonymous and non-synonymous substitutions demonstrated that both modes of evolutionary pressure, positive as well as negative selection, have acted. In addition, conservation of some β-defensin exons is demonstrated. Discrimination of certain β-defensin genes was possible only due to intron-specific differences. Therefore, sequence analyses restricted to the exons would result in underestimation of the number of β-defensin genes. Our study shows that for reconstruction of the phylogenetic history data of defensin introns are more appropriated. Comparisons among the amino acid sequences show moderate substitutions without changing the net charge of the mature peptides. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Human‐mediated introduction of introgressed deer across Wallace's line: Historical biogeography of Rusa unicolor and R. timorensis

In this study we compared the phylogeographic patterns of two Rusa species, Rusa unicolor and Rusa timorensis, in order to understand what drove and maintained differentiation between these two geographically and genetically close species and investigated the route of introduction of individuals to the islands outside of the Sunda Shelf. We analyzed full mitogenomes from 56 archival samples from the distribution areas of the two species and 18 microsatellite loci in a subset of 16 individuals to generate the phylogeographic patterns of both species. Bayesian inference with fossil calibration was used to estimate the age of each species and major divergence events. Our results indicated that the split between the two species took place during the Pleistocene, ~1.8 Mya, possibly driven by adaptations of R. timorensis to the drier climate found on Java compared to the other islands of Sundaland. Although both markers identified two well‐differentiated clades, there was a largely discrepant pattern between mitochondrial and nuclear markers. While nDNA separated the individuals into the two species, largely in agreement with their museum label, mtDNA revealed that all R. timorensis sampled to the east of the Sunda shelf carried haplotypes from R. unicolor and one Rusa unicolor from South Sumatra carried a R. timorensis haplotype. Our results show that hybridization occurred between these two sister species in Sundaland during the Late Pleistocene and resulted in human‐mediated introduction of hybrid descendants in all islands outside Sundaland.     

Whole genome sequencing and methylome analysis of the wild guinea pig

Background

DNA methylation is a heritable mechanism that acts in response to environmental changes, lifestyle and diseases by influencing gene expression in eukaryotes. Epigenetic studies of wild organisms are mandatory to understand their role in e.g. adaptational processes in the great variety of ecological niches. However, strategies to address those questions on a methylome scale are widely missing. In this study we present such a strategy and describe a whole genome sequence and methylome analysis of the wild guinea pig.

Results

We generated a full Wild guinea pig (Cavia aperea) genome sequence with enhanced coverage of methylated regions, benefiting from the available sequence of the domesticated relative Cavia porcellus. This new genome sequence was then used as reference to map the sequence reads of bisulfite treated Wild guinea pig sequencing libraries to investigate DNA-methylation patterns at nucleotide-specific level, by using our here described method, named ‘DNA-enrichment-bisulfite-sequencing’ (MEBS). The results achieved using MEBS matched those of standard methods in other mammalian model species. The technique is cost efficient, and incorporates both methylation enrichment results and a nucleotide-specific resolution even without a whole genome sequence available. Thus MEBS can be easily applied to extend methylation enrichment studies to a nucleotide-specific level.

Conclusions

The approach is suited to study methylomes of not yet sequenced mammals at single nucleotide resolution. The strategy is transferable to other mammalian species by applying the nuclear genome sequence of a close relative. It is therefore of interest for studies on a variety of wild species trying to answer evolutionary, adaptational, ecological or medical questions by epigenetic mechanisms.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1036) contains supplementary material, which is available to authorized users.     

Reproductive fitness in roe bucks (Capreolus capreolus): seasonal timing of testis function

The exact seasonal timing of normal testis function is a crucial precondition for the reproductive fitness of roe bucks and for successful breeding during rut in July–August. Production of spermatozoa and testosterone requires both endocrine regulation and local testicular control by autocrine/paracrine factors. These local control mechanisms include the action of several growth factors. Our short review assigns histological organization of roe deer testis to new data on the involvement of several growth factors in its regulation. The expression of growth factors is season-specific and cell-type-specific. This suggests its functional role in the complex interaction between germinative and somatic cells for the regulation of testis growth, spermatogenesis and function of hormone-producing cells. The authors dedicate this review to Prof. Dr. Christian Pitra who celebrates his 65th birthday in April 2006.     

Circannual changes in the expression of vascular endothelial growth factor in the testis of roe deer (Capreolus capreolus)

Adult roe deer males show seasonal cycles of testicular growth and involution. The exact timing of these cycles requires endocrine regulation and local testicular control by autocrine/paracrine factors. Recent findings suggest that the vascular endothelial growth factor (VEGF) might have effects on both vascular and germinative cells in testis. Thus, we studied the expression pattern of vascular endothelial growth factor (VEGF) in roe deer testis using quantitative RT-PCR. The strength of VEGF mRNA expression depended on season. It reached its highest level at the peak of spermatogenesis during the pre-rutting period and had its nadir at the end of the rut when involution already began. The results suggested that VEGF may directly affect the regulation of spermatogenesis but may not be involved predominantly in testicular microvasculature as initially expected.     

Diversity of Insects in Nature protected Areas (DINA): an interdisciplinary German research project

Biodiversity and Conservation - Insect declines and biodiversity loss have attracted much attention in recent years, but lack of comprehensive data, conflicting interests among stakeholders and...     

Low level of genetic variability in European bisons (Bison bonasus) from the Bialowieza National Park in Poland

Nuclear DNA markers (microsatellites) were used to screen the genetic variability in the European bison population of the Bialowieza National Park, Poland. The species is listed as endangered and the Bialowieza population is the largest one worldwide. Many other herds were founded by individuals from Bialowieza. Out of 18 microsatellites, nine were polymorphic, five were found to be homozygous, and four loci did not amplify. No significant deviation from Hardy–Weinberg-equilibrium (HWE) was observed, and the average number of alleles was 2.3 per locus. Thus, the European bison is characterized by a very low level of genetic diversity, most likely resulting from the population decline in the nineteenth century. Nevertheless, allelic variability derived from the nine polymorphic loci established in this study allowed to identify each individual by its genotypic profile. This data is valuable for conservation plans of this impressive species, especially for the control of breeding success in these animals.     

Nonconcordant evolutionary history of maternal and paternal lineages in Adriatic sturgeon

Although analyses of intraspecific variability are an important prerequisite for species identification assays, only a few studies have focused on population genetics and historical biogeography of sturgeon species. Here we present the first study on genetic variability of the last remaining Adriatic sturgeon, Acipenser naccarii, derived from mitochondrial and nuclear DNA. Our mitochondrial DNA analyses arranged individuals into three distinguished mitochondrial DNA haplogroups (Po1, Po2 and Buna). Two haplogroups (Po1 and Buna) were correlated to geographical distribution, whereas the third (Po2) was not. It was, however, very closely related to one lineage of its Ponto-Caspian sister species, A. gueldenstaedtii. The distribution of nuclear markers (microsatellites and amplified fragment length polymorphism) was strongly correlated to geographical distribution. An assignment test based on nuclear data placed no specimen of A. naccarii to A. gueldenstaedtii and vice versa. Therefore, the presence of gueldenstaedtii-like haplotypes within the Po population is either the result of a postglacial introgression or an ancestral polymorphism and does not indicate a hybrid population. The most valuable tool for forensic species identification purposes is one diagnostic deletion separating all A. naccarii from A. gueldenstaedtii. As both A. naccarii populations are genetically differentiated, stocking of sturgeon from the Po River in Italy into waters of the Buna River would jeopardize the genetic differences between both populations and should thus be avoided.