Marine Arakelyan

Mitochondrial phylogeography of European pond turtles ( Emys orbicularis , Emys trinacris ) – an update

Based on more than 1100 samples of Emys orbicularis and E. trinacris, data on mtDNA diversity and distribution of haplotypes are provided, including for the first time data for Armenia, Georgia, Iran, and the Volga, Ural and Turgay River Basins of Russia and Kazakhstan. Eight mitochondrial lineages comprising 51 individual haplotypes occur in E. orbicularis, a ninth lineage with five haplotypes corresponds to E. trinacris. A high diversity of distinct mtDNA lineages and haplotypes occurs in the south, in the regions where putative glacial refuges were located. More northerly parts of Europe and adjacent Asia, which were recolonized by E. orbicularis in the Holocene, display distinctly less variation; most refuges did not contribute to northern recolonizations. Also in certain southern European lineages a decrease of haplotype diversity is observed with increasing latitude, suggestive of Holocene range expansions on a smaller scale.

Variation of Mini- and Microsatellite DNA Markers in Populations of Parthenogenetic Rock Lizard Darevskia rostombekovi

Variation and clonal diversity in populations of the parthenogenetic rock lizard Darevskia rostombekovi was examined by means of multilocus DNA fingerprinting using mini- and microsatellite DNA markers M13, (GATA)4, and (TCC)50). The animals examined were shown to exhibit a clonally inherited, species-specific pattern of DNA markers (fingerprint profile) that is different from the species-specific patterns of parthenogenetic species D. dahli, D. armenica, and D. unisexualis. The mean intraspecific similarity indexS was 0.950 (0.003) for a sample of 19 animals from three isolated populations of North Armenia. This significantly differed from the estimate of this parameter for a sample of 21 animals including two individuals from mountainous, relict population from the vicinity of the Sevan Lake, which was equal to 0.875 (0.001). A comparison of DNA fingerprints showed differences between 21 individuals attaining 79 DNA fragments of 1801 mini- and microsatellite markers included in the analysis. The results obtained show that intraspecific variation in D. rostombekovi is higher than that in the previously studied parthenogenetic species D. dahli (S = 0.962) andD. unisexualis (S= 0.950) (P< 0.001). Taking into account that D. rostombekovi is considered monoclonal on the basis of allozyme data, the problem of clonal variability is discussed with regard to the evidence on nuclear DNA markers. It is suggested that the hybrid karyotype of D. rostombekovi, which is more unstable than that of D. dahli and D. unisexualis, generates a series of chromosomal rearrangements (mutations). This may lead to the appearance of a geographically isolated chromosomal race (clone) in the population inhabiting the southeastern coast of the Sevan Lake.

Hybrids of Darevskia valentini, D. armeniaca and D. unisexualis from a sympatric population in Armenia

A unique hybridization zone of rock lizard species of genus Darevskia, with diploid (2n), triploid (3n )a nd tetraploid (4n) individuals, occurs in the mountain steppe of central Armenia. Our long-term monitoring has showed an increasing birth rate of triploid hybrids in this mixed population. Among these hybrids, the frequency of captured males with fully developed reproductive systems and presumably fertile females also increased significantly. Consequently, intensive microevolution has taken place in this sympatric population. The morphological, ecological, cytological, histological, and parasitological characteristics of the hybrid Darevskia are presented here and compared with parental species.

Gene flow among deeply divergent mtDNA lineages of Testudo graeca (Linnaeus, 1758) in Transcaucasia

Using 10 polymorphic microsatellite loci and sequences of the mitochondrial cytochrome b gene, we examine gene flow in more than 100 spur-thighed tortoises from Transcaucasia and compare our findings with previously published AFLP and mtDNA data. While mtDNA sequences correspond to three deeply divergent clades and AFLP data suggest two distinct groups, microsatellite data indicate weak differentiation and extensive gene flow. We conclude that each marker system reflects a distinct episode in the evolutionary history of the Testudo graeca complex, corresponding to phases of vicariance and extensive gene flow. We hypothesize that the reciprocally monophyletic and deeply divergent mtDNA lineages reflect old vicariance events, while the conflicting nuclear markers are the legacy of younger episodes of extensive gene flow. The differentiation pattern found in the AFLP markers, with AFLP groups matching allopatrically or parapatrically distributed mitochondrial lineages, is likely to be older than the microsatellite differentiation, which could correspond to Holocene range expansions into Caucasian valleys and lower altitudes. Owing to the largely mutually exclusive distribution ranges of the deeply divergent mtDNA lineages, their identification with distinct subspecies is a reasonable and straightforward classification that facilitates communication and acknowledges the conspecifity of the involved evolutionary units. Consequently, Transcaucasia constitutes an intergradation zone of T. g. armeniaca, T. g. buxtoni and T. g. ibera.

Cryptic diversity and unexpected evolutionary patterns in the meadow lizard, Darevskia praticola (Eversmann, 1834)

Darevskia praticola differs from the other species of the genus in having a large but disjunct distribution, covering the Balkan and the Caucasus regions. Furthermore, most Darevskia species occupy saxicolous habitats, whereas D. praticola inhabits meadows and forest environments. Here we determine the phylogeographic and phylogenetic relationships of Darevskia praticola sensu lato and evaluate the current, morphology-based taxonomy. We sequenced two mtDNA genes (Cyt-b and ND4) and two nuclear loci (MC1R and RELN) for samples collected across the species range. Because our sequences amplified with the Cyt-b primers appear to represent a nuclear pseudogene we excluded this marker from the final analysis. Our results support monophyly of D. praticola and show its division into three clades. The first divergence, dated to the Late Pliocene, is between the Balkans and the Caucasus. The Caucasus lineage is further subdivided in a western Greater Caucasus and a Transcaucasia clade, likely due to subsequent differentiation during the Pleistocene. Our findings do not support the current taxonomic arrangement within D. praticola. The main geographic divergence likely happened due to a vicariance event associated with Plio-Pleistocene climatic and vegetation oscillations.

Parthenogenesis through the ice ages: A biogeographic analysis of Caucasian rock lizards (genus Darevskia).

Darevskia rock lizards include both sexual and parthenogenetic species, mostly distributed in the heterogeneous and ecologically diverse Caucasus. The parthenogenetic species originated via directional hybridogenesis, with only some of the sexual species known to serve as parentals. However, it remains unclear when and where these events happened and how many parental lineages were involved. A multilocus phylogeographic analysis was performed on the parthenogens D. unisexualis, D. bendimahiensis and D. uzzeli, and their putative maternal species D. raddei. Results show the parthenogenetic species all have relatively recent origins, approximately 200-70kyr ago, and at least three hybridization events were involved in their formation. Ecological niche models identify the region where hybridization events leading to the formation of D. unisexualis took place, namely in the northeast of the current distribution. Models also suggest that the sexual D. raddei might have undergone a habitat shift between the Last Interglacial and the Last Glacial Maximum.

Reticulate Evolution of the Rock Lizards: Meiotic Chromosome Dynamics and Spermatogenesis in Diploid and Triploid Males of the Genus Darevskia

Knowing whether triploid hybrids resulting from natural hybridization of parthenogenetic and bisexual species are fertile is crucial for understanding the mechanisms of reticulate evolution in rock lizards. Here, using males of the bisexual diploid rock lizard species Darevskia raddei nairensis and Darevskia valentini and a triploid hybrid male Darevskia unisexualis × Darevskia valentini, we performed karyotyping and comparative immunocytochemistry of chromosome synapsis and investigated the distribution of RAD51 and MLH1 foci in spread spermatocyte nuclei in meiotic prophase I. Three chromosome sets were found to occur in cell nuclei in the D. unisexualis × D. valentini hybrid, two originating from a parthenogenetic D. unisexualis female and one from the D. valentini male. Despite this distorted chromosome synapsis and incomplete double-strand breaks repair in meiotic prophase I, the number of mismatch repair foci in the triploid hybrid was enough to pass through both meiotic divisions. The defects in synapsis and repair did not arrest meiosis or spermatogenesis. Numerous abnormal mature spermatids were observed in the testes of the studied hybrid.

A standard photomap of the ovarian nurse cell chromosomes for the dominant malaria vector in Europe and Middle East Anopheles sacharovi

BackgroundAnopheles sacharovi is a dominant malaria vector species in South Europe and the Middle East which has a highly plastic behaviour at both adult and larval stages. Such plasticity has prevented this species from eradication by several anti-vector campaigns. The development of new genome-based strategies for vector control will benefit from genome sequencing and physical chromosome mapping of this mosquito. Although a cytogenetic photomap for chromosomes from salivary glands of An. sacharovi has been developed, no cytogenetic map suitable for physical genome mapping is available.MethodsMosquitoes for this study were collected at adult stage in animal shelters in Armenia. Polytene chromosome preparations were prepared from ovarian nurse cells. Fluorescent in situ hybridization (FISH) was performed using PCR amplified probes.ResultsThis study constructed a high-quality standard photomap for polytene chromosomes from ovarian nurse cells of An. sacharovi. Following the previous nomenclature, chromosomes were sub-divided into 39 numbered and 119 lettered sub-divisions. Chromosomal landmarks for the chromosome recognition were described. Using FISH, 4 PCR-amplified genic probes were mapped to the chromosomes. The positions of the probes demonstrated gene order reshuffling between An. sacharovi and Anopheles atroparvus which has not been seen cytologically. In addition, this study described specific chromosomal landmarks that can be used for the cytotaxonomic diagnostics of An. sacharovi based on the banding pattern of its polytene chromosomes.ConclusionsThis study constructed a high-quality standard photomap for ovarian nurse cell chromosomes of An. sacharovi and validated its utility for physical genome mapping. Based on the map, cytotaxonomic features for identification of An. sacharovi have been described. The cytogenetic map constructed in this study will assist in creating a chromosome-based genome assembly for this mosquito and in developing cytotaxonomic tools for identification of other species from the Maculipennis group.

The Geographic Distribution of Turtles in Armenia and the Nagorno-Karabakh Republic (Artsakh)

ABSTRACT The turtles of the Caucasus are poorly studied, and basic data on their geographic distribution in some Caucasian regions are poorly collated, hard to access, or entirely lacking. We present a comprehensive review of turtle localities in such a region, Armenia and Nagorno-Karabakh Republic (Artsakh), based on literature records, personal observations, and museum specimens. The compiled data for the 3 native species (Emys orbicularis, Mauremys caspica, and Testudo graeca) show that their distributions in this region are restricted to 3 areas of low elevation. In the northeast, turtles can be found in the Kura River drainage. In the south and west, turtles can be found in the Arax River drainage. The distributions of species within these areas vary, with E. orbicularis being the most restricted by elevation. We note that the 3 regions here form part of an important transect from the interior Arax drainage (in the Ararat region of western Armenia) to near its confluence with the Kura drainage (in southern Armenia and Nagorno-Karabakh) to the Kura drainage sensu stricto (in northeast Armenia). Future studies of turtles in these areas will play an important role in understanding the historical biogeography of Caucasian turtles.

Parthenogenetic Darevskia lizards mate frequently if they have the chance: a quantitative analysis of copulation marks in a sympatric zone

Several Caucasian rock lizards of the genus Darevskia of hybrid origin are known to reproduce parthenogenetically. Local communities can be composed exclusively of parthenogens, though syntopy with bisexual members of the genus may occur. In some localities, reproduction between bisexual and parthenogenetic Darevskia has been previously reported based on lizard intermediate morphology and karyology (3n, 4n). However, the frequency of such heterospecific matings remains unknown. We indirectly quantified the reproductive interactions through the inspection of copulation marks in females in a mixed Darevskia community from Kuchak (Armenia) composed of two hybrid parthenogens (D. armeniaca and D. unisexualis), one bisexual species (D. valentini) and their putative backcrosses. A total of 139 adults were randomly collected and photographed. Females were later measured (SVL), inspected for inguinal marks and ranked from 0 (no scars) to 3 (≥ 3 scars). The lizard species and ploidy determination was ensured by a parallel microsatellite analysis. Sex-ratio in the community was extremely biased due to the high abundance of parthenogenetic females. All female types displayed copulation marks with frequencies varying from 80% in D. valentini to 64% in D. armeniaca. Remarkably, 7 out of 11 (64%) backcross females also showed marks. In the most abundant D. armeniaca, the prevalence and intensity of copulation marks increased with body size, just as predicted for polygynous female lacertids. These results indicate that copulation between parthenogenetic and bisexual species in Darevskia mixed communities is common and driven by sexual selection, thus reinforcing previous suggestions of reproductive interaction in syntopy. Evolutionary implications of these findings are discussed.