Population genetic studies in the Balkans. I. Serum proteins

Within a study of the genetics of Southeastern European populations seven serum protein polymorphisms (AMY2, BF, C3, CP, GC, HPA, TF) were examined in three samples of Aromuns (Albania: the village of Andon Poci, province Gjirocaster, Republic of Macedonia: Stip region, Romania: the village Kogalniceanu, province Dobruja) and four reference samples (Albanians: Tirana, Romanians: Constanta and Ploiesti as well as Greeks (Northeastern Greece)). The Aromun samples from Albania and Romania form one separate cluster and the reference samples together with the Aromuns from Macedonia (Stip region) form a second one.     

Genetic studies in south Balkan populations

Within a study of the genetics of Balkan populations, four DNA-STR systems and 19 classical markers were examined in seven samples: Romanians (two groups), Albanians, Greeks and Aromuns (three groups). The results for the DNA-STR systems have been compared with data from the literature. The results show four clear separated groups: sub-Saharan black populations, North-African, Japanese and European populations. The large Balkan populations, except the Greek sample, are genetically more homogenous than the Aromun populations. A second Neighbor-joining tree based on all 23 analyzed systems, show a particular trend of the Aromun groups, which indicates a particular genetic structure.     

Phenotype and allele frequencies of some serum protein polymorphisms in populations of the Balkans

Within a study of the genetics of Southeastern European populations seven serum protein polymorphisms (AMY2, BF, C3, CP, GC, HPA, TF) were examined in two samples of Aromuns and one reference sample (Musequiar-Aromuns from Dukasi in Albania, Moskopolian-Aromuns from Krusevo, Republic of Macedonia, and Macedonians from Skopje). The neighbor joining tree as well as the principal component analysis show results which do not correspond well to the geographic and historic background. This indicates that in the present case the serum protein polymorphisms give no clearly defined information about the relationships between the Balkan populations and to the origin of Aromuns.     

Dermatoglyphic studies in eastern and south-eastern Europe

In nine population samples from Bulgaria, Croatia, Greece, Hungary, Republic of Macedonia (Skopje and Aromuns from Stip region), Romania, Serbia and Slovakia 12 dermatoglyphic variables have been studied. There are distinct differences between the populations and between males and females. The Macedonian Aromuns are clearly separated from the other populations.     

Allele frequencies of D21S11, FGA, TH01, and VWA in populations of the Balkans

This study is part of an extensive investigation of the genetic relationship between Balkan populations, especially the Aromuns. Allele frequencies of four STRs (D21S11, FGA, TH01, VWA) from Macedonians (Skopje), Gramostian Aromuns from the Stip region (Macedonia), Moskopolian Aromuns from Krusevo (Macedonia), and Musequiar Aromuns from Dukasi (Albania) are presented.     

DNA-PCR systems TH01 and VWA31: population data from Albania and Romania

The microsatellite systems TH01 and VWA31 have been typed and analyzed for three populations from Albania (city population of Tirana, and Aromuns from a small-sized village, named Andon Poci, near Gjirokaster) and Romania (city population of Constanta). The genetic data have been compared with other, particularly South-Eastern European population.     

Population genetic studies of the pancreatic amylase (AMY2, E.C. 3.2.1.1) in Bulgaria

The pancreatic amylase (AMY2, E.C. 3.2.1.1) polymorphism has been studied in 2346 individuals from south-central and south-eastern Bulgaria. The allele frequencies have been determined as AMY2*1 = 0.9520 and AMY2*2 = 0.0480. The neighbor joining tree of seven subpopulations revealed only small genetic distances. Compared with other populations, the Bulgarian sample clustered with samples from Romania, Hungary, Germany and Switzerland, with larger distances to Albania, Greece and Macedonia.     

Isonymy, consanguinity and repeated pairs of surnames in Aromun populations

The Aromuns represent a small and almost unknown people that live scattered over the Balkan Peninsula. Due to their language, that is very similar to classical Latin, they are in a special position. The Aromuns settled only in more recent times. Until now they lived as shepherds, as caravan guides and merchants and lead a semi-nomadic life. We are currently carrying out studies to determine the genetic structure of this population. To facilitate the interpretation of these data, we are also trying to obtain other important parameters that pertain to migration processes and the genealogical structure of this populations. The data arise from three areas in Albania, the Republic of Macedonia and Romania. The inbreeding coefficient and the proportion of repeated pairs of surnames was calculated through the use of genealogies and the isonymy method. The difference between these three populations are due primarily to confounding by selection of mates and family composition.     

RFLP diversity and relationships among traditional European maize populations

Given the large extent of hybrid cultivation, the importance of conserving the diversity of crop genetic resources has given birth to numerous collections of old races. In the present paper, we conduct a molecular characterisation of a large collection of 488 European maize populations using the bulk RFLP analysis. The analysis of 23 RFLP loci showed a high allelic richness of 11.5 alleles per locus. Populations from eastern Europe (Poland, Austria, Germany, etc.) showed the lowest genetic diversity, a lower number of unique alleles and a higher percentage of fixed loci than populations from southern Europe. In fact, genetic diversity appeared higher in Southern regions where the first maize populations are thought to have been introduced. Molecular classification based on Rogers' distance (i.e. alleles frequencies) allowed us to distinguish three main clusters which were highly consistent with geographic origins. A Northeastern cluster grouped together early or intermediate populations from Northeastern countries and the Balkans, a southeastern cluster joined late and partially dent populations from Greece and Italy, and, a southwestern cluster was made up of early flint populations from northern Spain, Portugal and the Pyrenees. A correlation between allelic frequencies at some loci and latitude and/or longitude was observed. Such tendencies may reflect the direction of gene flow between different races of maize: for instance, North American (Northern flint) and Caribbean populations were introduced, respectively, to northern and southern Europe, in the past.     

Human population genetic structure detected by pain-related mu opioid receptor gene polymorphisms

Several single nucleotide polymorphisms (SNPs) in the Mu Opioid Receptor gene (OPRM1) have been identified and associated with a wide variety of clinical phenotypes related both to pain sensitivity and analgesic requirements. The A118G and other potentially functional OPRM1 SNPs show significant differences in their allele distributions among populations. However, they have not been properly addressed in a population genetic analysis. Population stratification could lead to erroneous conclusions when they are not taken into account in association studies. The aim of our study was to analyze OPRM1 SNP variability by comparing population samples of the International Hap Map database and to analyze a new population sample from the city of Corrientes, Argentina. The results confirm that OPRM1 SNP variability differs among human populations and displays a clear ancestry genetic structure, with three population clusters: Africa, Asia, and Europe-America.     

Assessment of genetic relationships among native and introduced Himalayan balsam (Impatiens glandulifera) plants based on genome profiling

We conducted genomic characterization based on SNP and SilicoDArT markers on the invasive Himalayan balsam (Impatiens glandulifera) plants originating from native and non‐native regions of their distribution. When genetic relationships were explored by PCoA using SNP and SilicoDArT marker data, the first, second, and third principal coordinates explained altogether 37.4% and 31.0% of the variability, respectively. Samples from the UK, Canada, and Pakistan were grouped together, while Indian plants were clearly distinct based on SNP markers but relatively close to the UK–Canada–Pakistan group based on SilicoDArT markers. Constructed trees differentiated individuals into clusters resembling the PCoA patterns. The Bayesian BAPS analysis performed for the SNP data revealed that the individuals were distributed in seven clusters, representing samples from each of the four Finnish populations, India, Pakistan, and the combination of the UK and Canada. Similar clustering was visible in the UPGMA tree. The Indian cluster did not display any ancestral gene flow with the others, while the Pakistani cluster showed ancestral gene flow only with the combined UK and Canada cluster. Furthermore, the latter cluster displayed ancestral gene flow with the Finnish populations varying from 0% to 3.1%. The BAPS analyses conducted for the SilicoDArT data differ slightly: The individuals were distributed in nine clusters, and the Indian cluster exhibited ancestral gene flow with the mixed cluster including Canadian, Pakistani, and UK samples, and one Finnish sample. The AMOVA showed that 45% and 26% of variation was present among the I. glandulifera groups/populations and the rest within them based on SNP and SilicoDArT markers, respectively. The Bayesian BAPS analyses and the gene flow networks were the most informative tools for resolving relationships among native and introduced plants. It is notable that the small sample sizes for non‐Finnish plant materials may affect the accuracy of the gene flow and other estimates.     

Nested clade and phylogeographic analyses of the chub, Leuciscus cephalus (Teleostei, cyprinidae), in Greece: implications for Balkan Peninsula biogeography

Phylogenetic relationships among Greek populations of the chub, Leuciscus cephalus, were investigated using 600 bp of the cytochrome b gene. The aim of this study was to test the assumption that the main difference in ichthyological composition between both sides of the Balkan Peninsula is directly linked to differences in the dispersion mechanisms used by fish in order to extend their distribution range. Phylogenetic and nested clade analyses clearly showed that populations in Greece are significantly differentiated. Greek populations were found to descend from three lineages in three geographical provinces: Western, Central, and Eastern Greece. The chub reached Western Greece at the beginning of the Pleistocene and Eastern Greece during the mid-Pleistocene. Chub dispersion occurred mainly by river confluence due to sea level lowering and river capture in Western Greece and sea dispersal with low-salinity conditions within the Aegean Sea in Eastern Greece. However, in Central Greece, the original mtDNA lineage has presumably been lost owing to a genetic introgression following a second invasion from the Danube during the final stage of the last glaciation. This study provides new elements for a better understanding of the composition of the contemporary ichthyofauna in Greece and highlights possible evolutionary mechanisms responsible for the high endemism rate in the Western Greek biogeographic province.     

An Application of Serially Balanced Designs for the Study of Known Taste Samples with the α-ASTREE Electronic Tongue

The α-ASTREE e-Tongue instrument uses seven sensors to characterize taste signals associated with a liquid sample. The instrument was used to study eight test preparations (comprised of a blank, four preparations corresponding to four known tastes and Sodium Topiramate in three concentrations known to have a bitter taste) and eight washes. Serially balanced residual effects designs were used to order the samples to estimate residual and main effects. The design provided for eight repeated measurements per test preparation. The experimental results suggested the following: (1) The seven sensors can be separated into three groups according to the ability to discriminate test preparations, and three of the sensors contributed little or no information. Further investigation suggested the lack of differentiability might be due to the age of the sensors. (2) The sensors discriminated known tastes from blank. The residual effect due to test preparations might appear after repeated usage. (3) Exploratory principal component analysis of the data indicated that nearly 90% of the total variability across the seven sensors could be explained by a single principal component. (4) The four standard taste preparations did not correspond to orthogonal dimensions in the principal component axes. (5) The three Sodium Topiramate test preparations could neither be associated with the corresponding known bitter taste sample nor could the three doses be shown to follow a quantitative dose-response relationship on the e-Tongue measurement scale. The practical interpretation of the results of the statistical analysis indicates only poor discriminative ability of the e-Tongue to distinguish clearly between increasing concentrations of a known bitter compound such as Sodium Topiramate. No apparent linear relationship could be discerned over increasing concentrations that would allow the quantification of bitterness.KEY WORDS: cluster analysis, electronic tongue, principal component analysis, residual effects designs, serially balanced design     

Further evidence supporting Morocco as a centre of origin of barley

 Thirty-five populations of H. spontaneum from nine countries, encompassing almost all the known range of distribution of the species, Afghanistan, Crete (Greece), Cyprus, Iran, Iraq, Israel, Libya, Morocco and Turkey, were studied utilizing RFLP markers (21 probes with three restriction enzymes) distributed across all seven barley chromosomes in an attempt to unveil the genetic dissimilarities existing among them. UPGMA clustering, based on the Nei and Li (1979) similarity coefficient, produced a dendrogram where three clusters could be defined: two with a clear geographical distinction (Morocco and Cyprus) and another one grouping all the Asian/Middle Eastern populations, except for an accession from Iran that clustered separately. These results confirm our previous work and suggest that barley domestication could also have taken place outside the Fertile Crescent, particularly in Morocco. Received: 10 October 1998 / Accepted: 27 October 1998     

Genetic survey of shallow populations of the Mediterranean red coral [Corallium rubrum (Linnaeus, 1758)]: new insights into evolutionary processes shaping nuclear diversity and implications for conservation

Combined action from over‐harvesting and recent mass mortality events potentially linked to ongoing climate changes has led to new concerns for the conservation of shallow populations (5–60 m) of Corallium rubrum, an octocorallian that is mainly found in the Mediterranean Sea. The present study was designed to analyse population structure and relationships at different spatial scales (from 10s of meters to 100s of kilometres) with a focus on dispersal pattern. We also performed the first analysis of the distribution of genetic diversity using a comparative approach between regional‐clusters and samples. Forty populations dwelling in four distinct regions between 14 and 60 m in depth were genotyped using 10 microsatellites. Our main results indicate (i) a generalized pair‐sample differentiation combined with a weak structure between regional‐clusters; (ii) the occurrence of isolation by distance at the global scale, but also within two of the three analysed regional‐clusters; (iii) a high level of genetic diversity over the surveyed area with a heterogeneous distribution from regional‐cluster to sample levels. The evolutionary consequences of these results are discussed and their management implications are provided.     

Fagus sylvatica forest vegetation in Greece: Syntaxonomy and gradient analysis

Abstract. This is the first comprehensive study on Fagus sylvatica s.l. forest communities in Greece. It is based on the multivariate analysis of more than 1100 relevés throughout the distribution area of Fagus in Greece, 60 % of which have been sampled within the last 15 years. The data were compiled from all relevant literature and from unpublished field studies. Prior to data analysis, taxonomic and floristic inaccuracies in older literature have been corrected, and some taxa aggregated. Status and distribution of Fagus in Greece, as well as historical and silvicultural aspects, are outlined. Classification resulted in 16 vegetation types defined by species composition, of which 10 units are ranked as association (of which two were described as new) or community, with six subcommunities included. Synonyms from the literature are given to all types. The floristic structure of the data set is displayed by a synoptic table and by DCA ordination. Community differentiation follows a complex edaphic‐climatic‐phytogeographic pattern. Apart from Fagus sylvatica ssp. orientalis forests of warm and dry habitats and low to medium altitudes which are related to deciduous Quercus forests, four Fagion community groups are distinguished which represent different habitat types: (1) cool, humid, mesotrophic; (2) cool, acidic; (3) high altitude, calcareous; (4) medium altitude, moderately warm and dry. The communities may be assigned to three regional suballiances, viz. (a) Doronico orientalis‐Fagenion at medium altitudes of North‐Central and East‐Central Greece; (b) Doronico columnae‐Fagenion in the far north and at high altitudes southward to East‐Central Greece; (c) Geranio striati‐Fagenion in the Northwest. Various types of thermophilous beech forest are assigned to (a) and (c), acidic forest to (b), and mesic Fagus forest to (b) and (c).     

Genetic diversity of sharpnose mullet <Emphasis Type="Italic">Liza saliens</Emphasis> introduced in southern Caspian Sea in comparison with one native Aegean Sea population

Sharpnose mullet, Liza saliens (Risso, 1810) is one of the valuable species in Caspian shoreline and it was first introduced to Caspian Sea from Black Sea between 1930 and 1934. In the present study, we used six microsatellite markers to obtain genetic information for L. saliens from four localities of southern Caspian Sea and for one native population from Greece coastline (northern Aegean Sea). Results showed lower number of alleles (Na = 5.7–6) and higher observed heterozygosity (Ho = 0.74–0.83) in Caspian Sea samples, related to the native Aegean population (Na = 8, Ho = 0.68). Significant deviations from Hardy-Weinberg expectations were detected in 19 out of 28 tests. Low genetic differentiation (provided Fst values) among samples was detected, but Behshahr sample was significantly differentiated from all the others. The UPGMA tree revealed three major clusters: the Behshahr sample and Greek population were clustered in two different clades, and the remaining three samples formed another cluster. The low genetic variation and spatial differentiation among Caspian Sea samples may be justified by the recent establishment.     

Genetic diversity and morphological variability in the Balkan endemic Campanula secundiflora s.l. (Campanulaceae)

We reconstructed the relationships among populations of Campanula secundiflora s.l. and closely related and geographically close populations of C. austroadriatica and C. versicolor. Based on analyses of microsatellite DNA data, the investigated populations have high overall genetic diversity and abundant allelic variation over seven investigated loci. Bayesian model‐based clustering identified four clearly differentiated genetic groups of populations. The genetic differentiation was reflected by morphological differentiation, allowing us to propose a new taxonomic treatment of the constituents of C. secundiflora s.l. The populations distributed in south‐western Serbia and north‐eastern Montenegro were included in C. secundiflora. A new species, C ampanula montenegrina sp. nov. , distributed in the continental part of Montenegro and the northern part of Albania is described. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 64–88.     

Mitochondrial DNA variations in Russian and Belorussian populations

The sequence of the first hypervariable segment (HVS-I) of mitochondrial DNA (mtDNA) was determined in 251 individuals from three eastern Slavonic populations, two Russian and one Belorussian. Within HVS-I, 78 polymorphic positions were revealed. Within-population diversity of HVS-I varies slightly among three samples; its estimates do not differ strongly from those for European populations. Haplotype diversity for three populations calculated in this study is 0.949; mean pairwise differences estimate is 3.59. To assign mtDNA sequences to major phylogenetic clusters, haplogroup-specific restriction polymorphisms were selectively typed in most samples. The haplogroup distribution in the total Eastern Slavonic sample is similar to that reported for the European sample. However, the separate consideration of three Slavonic samples reveals the complicated structure of the mitochondrial gene pool in the Eastern European area. Data of this study support the proposed model of the origin of modern Eastern Slavs, which implies the admixture of ancient Slavonic tribes with pre-Slavonic populations of Eastern Europe. These data should contribute to general studies of mitochondrial DNA variations in Europe.     

Comparing the genetic structure of codling moth Cydia pomonella (L.) from Greece and France: long distance gene-flow in a sedentary pest species

Codling moth Cydia pomonella L. (Lepidoptera: Tortricidae) is the most important insect pest of apple production in Europe. Despite the economic importance of this pest, there is not information about the genetic structure of its population in Greece and the patterns of gene-flow which might affect the success of control programs. In this study, we analysed nine samples from apple, pear and walnut from various regions of mainland Greece using 11 microsatellite loci. Six samples from the aforementioned hosts from southern France were also examined for comparison. Bayesian clustering and genetic distance analyses separated the codling moth samples in two genetic clusters. The first cluster consisted mainly of the individuals from Greece, and the second of those from France, although admixture and miss-classified individuals were also observed. The low genetic differentiation among samples within each country was also revealed by F(ST) statistics (0.009 among Greek samples and 0.0150 among French samples compared to 0.050 global value among all samples and 0.032 the mean of the pair-wise values between the two countries). These F(ST) values suggest little structuring at large geographical scales in agreement with previous published studies. The host species and local factors (climatic conditions, topography, pest control programs) did not affect the genetic structure of codling moth populations within each country. The results are discussed in relation to human-made activities that promote gene-flow even at large geographic distances. Possible factors for the genetic differentiation between the two genetic clusters are also discussed.