A molecular basis for natural selection at the timeless locus in Drosophila melanogaster

Diapause is a protective response to unfavorable environments that results in a suspension of insect development and is most often associated with the onset of winter. The ls-tim mutation in the Drosophila melanogaster clock gene timeless has spread in Europe over the past 10,000 years, possibly because it enhances diapause. We show that the mutant allele attenuates the photosensitivity of the circadian clock and causes decreased dimerization of the mutant TIMELESS protein isoform to CRYPTOCHROME, the circadian photoreceptor. This interaction results in a more stable TIMELESS product. These findings reveal a molecular link between diapause and circadian photoreception.     

Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases

Nineteen dogs from Greece with chronic ehrlichiosis were studied. The dogs exhibited bicytopenia or pancytopenia, bone marrow hypoplasia, seroreactivity to Ehrlichia canis (E. canis) antigens, and had no history of drug or radiation exposure. Anorexia, depression, severe bleeding tendencies, hypoalbuminemia, and increased serum alanine aminotransferase activity were also hallmarks of the disease. All these animals eventually died, irrespective of the treatment applied. Some dogs were also serologically positive for Rickettsia conorii, Leishmania infantum (L. infantum), and Bartonella vinsonii subspp. berkhoffii. Polymerase chain reaction testing of bone marrow samples revealed E. canis, Anaplasma phagocytophilia, Anaplasma platys, and L. infantum in some dogs. Concurrent infections did not appear to substantially influence the clinical course and final outcome of the chronic canine ehrlichiosis.     

Development of a national crop wild relative conservation strategy for Cyprus

Growing research interest in crop wild relatives (CWR) has highlighted their value for crop improvement, particularly to mitigate the impact of climate change and contribute to global food security. As most conservation activities are implemented at national level there is a requirement for each country to develop and implement a national CWR conservation strategy. This will contribute to regional and global CWR conservation actions and so help achieve CBD Aichi Targets 13 of improved genetic conservation of socio-economically important taxa. Cyprus has historically lacked systematic CWR conservation but is rich in CWR diversity being located in the eastern Mediterranean Vavilov centre and abutting the Fertile Crescent. Therefore a more coherent approach to CWR conservation is desirable. From an initial checklist of 1,722 Cypriot CWR taxa, 178 CWR were prioritized and eco geographic data was collated and analysed using GIS techniques. The Troodos and Pafos mountains are CWR hotspots and ten priority sites containing the genetic diversity of 74.7 % of priority CWR taxa were identified. These will form the basis of a national network of in situ genetic reserves and 78 priority CWR taxa that have no accessions in the national Cyprus gene bank are highlighted for collection and ex situ storage.     

Inhibitory activity on amyloid-beta aggregation and antioxidant properties of Crocus sativus stigmas extract and its crocin constituents

Crocus sativus stigmas are one of the widely known spices (saffron) and consist of unusually polar carotenoids. Alzheimer's disease is characterized pathologically by deposition of amyloid beta-peptide (Abeta) fibrils. Oxidation is thought to promote Abeta fibril formation and deposition. To identify agents inhibiting the pathogenesis of Alzheimer's disease, we examined in vitro the antioxidant properties of extract of C. sativus stigmas and its effect on Abeta(1-40) fibrillogenesis. The antioxidant properties were determined by measuring the ferric-reducing antioxidant power and Trolox-equivalent antioxidant capacity, while its effects on Abeta-aggregation and fibrillogenesis were studied by thioflavine T-based fluorescence assay and by DNA binding shift assay. The water:methanol (50:50, v/v) extract of C. sativus stigmas possesses good antioxidant properties, higher than those of tomatoes and carrots, and inhibited Abeta fibrillogenesis in a concentration and time-dependent manner. The main carotenoid constituent, trans-crocin-4, the digentibiosyl ester of crocetin, inhibited Abeta fibrillogenesis at lower concentrations than dimethylcrocetin, revealing that the action of the carotenoid is enhanced by the presence of the sugars. Our findings suggest the possible use of C. sativus stigma constituents for inhibition of aggregation and deposition of Abeta in the human brain.     

Transmission and pathogenicity of encephalomyocarditis virus (EMCV) among rats

Due to the probable role played by rodents as a reservoir for the transmission of the EMC virus to pigs, the experiment reported here was performed in order to assess the transmission rate of EMCV within a rat population. Twenty-five eight-week-old Wistar rats housed in individual plastic cages were experimentally infected either with a Greek myocardial EMCV strain (5 rats with a 0.2 x 10(6) TCID50 dose per rat and 10 rats with a 0.5 x 10(4.5) TCID50 dose per rat, oronasally) or a Belgian myocardial EMCV strain (10 rats with a 0.5 x 10(4.5) TCID50 dose per rat, oronasally). Two to five days later, each inoculated rat was moved to a new clean cage and coupled with a contact rat to compare the pathogenicity of the two strains and to estimate the basic reproduction ratio R0, indicating the level of EMCV transmission. During the experiments, faecal virus excretion was measured as well as the serological response against EMCV. After euthanasia, virus isolation was attempted from different rat tissues. Neither strains produced mortality, nor clinical signs and only low titres of neutralising antibodies were found. All contact rats, however, were infected and the virus was isolated from their faeces and from various tissues. Both 10-pair experiments revealed a point estimate for the R0 of infinity (95%-CI for both the Greek and Belgian EMCV strains = 4.48 - infinity), as did the 5-pair experiment with a higher dose of the Greek strain (95%-CI = 1.83 - infinity). Combining the results from the two 10-pair experiments resulted in an estimate for R0 of infinity (95%-CI: 9.87 - infinity). These results indicate that the EMC virus can spread very easily within a rat population by horizontal rat-to-rat transmission (R0 > 1).     

Natural selection favors a newly derived timeless allele in Drosophila melanogaster

Circadian and other natural clock-like endogenous rhythms may have evolved to anticipate regular temporal changes in the environment. We report that a mutation in the circadian clock gene timeless in Drosophila melanogaster has arisen and spread by natural selection relatively recently in Europe. We found that, when introduced into different genetic backgrounds, natural and artificial alleles of the timeless gene affect the incidence of diapause in response to changes in light and temperature. The natural mutant allele alters an important life history trait that may enhance the fly's adaptation to seasonal conditions.     

The geographic origin of old Douglas-fir stands growing in Central Europe

Since the nineteenth century, Douglas-fir seed sources have been widely used for establishment of forest stands outside its natural distribution range. In Europe, some of these old Douglas-fir stands are registered as seed stands and provide seed sources for nurseries, although it is unclear from which region in North America they originate. In recent years, the interest in planting Douglas-fir has increased substantially because the species is seen as a potential adaptation option to climate change. This makes the assignment of European Douglas-fir stands of unknown seed origin to their geographic origin in North America increasingly important, because the genetic quality of these plantations must be guaranteed. In this study, we use 13 nuSSR loci to investigate the origin of 67 Austrian and German Douglas-fir stands of unknown origin. We performed a hierarchical Bayesian cluster analysis using 38 native Douglas-fir populations. The resulting clusters are used as reference populations to assign the 67 Central European Douglas-fir stands from Austria and Germany planted more than 80 years ago. Our results suggest that the majority of our investigated Douglas-fir stands come from central Washington (USA), the recommended seed zones for Central Europe. Some stands were located outside the suggested area, e.g. central Oregon and Santa Fe (New Mexico). The accuracy assessment of our approach revealed the best performance for the highest hierarchical level, e.g. assigning populations either to the coastal or the Rocky Mountain variety. As expected, the uncertainty increases with decreasing hierarchical level. The final assessment, if an admixture of seed sources within the European Douglas-fir stands is evident suggests that 23 of the Douglas-fir stands show an admixture which was not detected in our Douglas-fir reference populations growing in the natural distribution range.     

PrP gene polymorphisms in Cyprus goats and their association with resistance or susceptibility to natural scrapie

In contrast to scrapie in sheep, the genetic basis of susceptibility to scrapie in goats is not well understood. To study the association of prion protein (PrP) alleles with susceptibility to scrapie in goats in Cyprus, the coding sequence of the caprine PrP gene was determined in 717 goats, including 218 scrapie positive animals. Several novel polymorphisms were detected, such as a novel octarepeat variant and a stop codon mutation. Amino acids at codons 146 and 154 were associated with susceptibility to goat scrapie. Animals heterozygous for serine (S) and aspartate (D) at codon 146 were significantly under-represented in scrapie positive animals and no positive animals were found that were homozygous for these amino acids at codon 146. These results might provide the basis for genetic control of scrapie in Cypriot goats.     

Interfertile oaks in an island environment: I. High nuclear genetic differentiation and high degree of chloroplast DNA sharing between <Emphasis Type="Italic">Q. alnifolia</Emphasis> and <Emphasis Type="Italic">Q. coccifera</Emphasis> in Cyprus. A multipopulation study

The evergreen Quercus alnifolia and Q. coccifera form the only interfertile pair of oak species growing in Cyprus. Hybridization between the two species has already been observed and studied morphologically. However, little evidence exists about the extent of genetic introgression. In the present study, we aimed to study the effects of introgressive hybridization mutually on both chloroplast and nuclear genomes. We sampled both pure and mixed populations of Q. alnifolia and Q. coccifera from several locations across their distribution area in Cyprus. We analyzed the genetic variation within and between species by conducting analysis of molecular variance (AMOVA) based on nuclear microsatellites. Population genetic structure and levels of admixture were studied by means of a Bayesian analysis (STRUCTURE simulation analysis). Chloroplast DNA microsatellites were used for a spatial analysis of genetic barriers. The main part of the nuclear genetic variation was explained by partition into species groups. High interspecific differentiation and low admixture of nuclear genomes, both in pure and mixed populations, support limited genetic introgression between Q. alnifolia and Q. coccifera in Cyprus. On the contrary, chloroplast DNA haplotypes were shared between the species and were locally structured suggesting cytoplasmic introgression. Occasional hybridization events followed by backcrossings with both parental species might lead to this pattern of genetic differentiation.     

Interfertile oaks in an island environment. II. Limited hybridization between <Emphasis Type="Italic">Quercus alnifolia</Emphasis> Poech and <Emphasis Type="Italic">Q. coccifera</Emphasis> L. in a mixed stand

Hybridization and introgression between Quercus alnifolia Poech and Q. coccifera L. is studied by analyzing morphological traits, nuclear and chloroplast DNA markers. The study site is a mixed stand on Troodos Mountains (Cyprus), and the analyzed material includes both adult trees and progenies of specific mother trees. Multivariate analysis of morphological traits shows that the two species can be well distinguished using simple leaf morphometric parameters. A lower genetic diversity in Q. alnifolia than in Q. coccifera and a high interspecific differentiation between the two species are supported by an analysis of nuclear and chloroplast microsatellites. The intermediacy of the four designated hybrids is verified by both leaf morphometric and genetic data. Analysis of progeny arrays provides evidence that interspecific crossings are rare. This finding is further supported by limited introgression of chloroplast genomes. Reproductive barriers (e.g. asynchronous phenology, postzygotic incompatibilities) might account for this result. A directionality of interspecific gene flow is indicated by a genetic assignment analysis of effective pollen clouds with Q. alnifolia acting as pollen donor. Differences in flowering phenology and species distribution in the stand may have influenced the direction of gene flow and the genetic differentiation among effective pollen clouds of different mother trees within species.