We have assessed the phylogenetic status of the Leishmania genome project Friedlin reference strain by MLEE and multiprimer RAPD including a set of 9 stocks representative of the main Leishmania species and of the whole genetic diversity of the Leishmania genus. To our knowledge, the detailed genetic characterization of the Friedlin strain has never been published before. As previously recorded (Tibayrenc et al. 1993), MLEE and RAPD data gave congruent phylogenetic results. The Friedlin reference strain was definitely attributed to Leishmania (Leishmania) major Yakimoff et Schokhor, 1914. Five specific RAPD patterns made it possible to distinguish between the Friedlin strain and the 2 other L. (L.) major stocks included in the study. Various specific MLEE and RAPD characters permitted to distinguish between the Leishmania species included in the study. All these characters are usable to detect accidental laboratory mix-ups involving the Friedlin reference strain. In confirmation with previous studies involving a more limited set of genetic markers, the general genetic diversity of the Leishmania genus proved to be considerable. It must be made clear that only one strain cannot be considered as representative of the whole genetic variability of the genus Leishmania. In the future, it is therefore advisable to complement the results obtained in the framework of the Leishmania genome project with data from other strains that should be selected on a criterion of important genetic differences with the Friedlin strain.
The Siah Mahi, Capoeta capoeta gracilis, is a widely distributed taxon in the south Caspian Sea basin (north of Iran) that has not yet been surveyed at the intra-specific level. Besides its ecological significance, this species is important for inland water fishing, aquaculture, sport fishing and zoogeographical studies. Here, we describe patterns of morphological and molecular differentiation among six populations of C. c. gracilis located in the Aras, Sefidrud, Shirud, Tonekãbon, Harãz and Gorgãnrud river systems in Iran. Univariate analysis of variance revealed significant differences between means of the six samples for 56 out of 60 morphometric ratios. The first two ordination axes of the mean ratios of the six samples (59.11% of total variation) separated all of the samples from each other, although not necessarily with any clear geographic pattern. The overall assignment of individuals into their original groups was high (88.6%). The proportion of individuals correctly classified into their original groups was 92.5%, 78.9%, 96%, 89.7%, 93.3% and 83.3% for Aras, Gorgãnrud, Sefidrud, Shirud, Tonekãbon and Harãz, respectively. Molecular clustering also grouped individuals of each sample either in the same cluster or the same sub-cluster. There was a large-scale congruence between results of the morphological and molecular analyses.
RAPD-PCR method is now widely employed in genetic research and is a powerful asset when trying to identify species. In the present work, RAPD genetic markers were obtained for distinguishing between three species of the genus Gobio that show substantial morphological resemblance. Different profiles were observed: six species-specific markers were detected for G. gobio, four for G. kesslerii and nine for G. uranoscopus. These markers were reproducible, dependable and very helpful in differentiating between these species.
The random amplified polymorphic DNA (RAPD) method was applied to eight Gobio gobio populations living in the rivers of the Czech Republic and Slovakia. The application of seven RAPD primers yielded eight fingerprint characteristics for the populations examined. Forty diagnostic markers have been identified, which can reliably identify the populations under study. Intrapopulation diversity varied between 0.26 and 0.38. A phenogram documented the close agreement of the particular populations with the geographic pertinence of their localities to the different sea basins.
Crested newts (Triturus cristatus superspecies) are a group of closely related species with parapatric distributions that are likely to interbreed where their ranges meet. Coexistence of three species of the complex (Triturus cristatus, T. dobrogicus and T. carnifex) has been recently confirmed in central Europe. In this study we aim to elucidate the distribution of crested newts in contact zones in the Czech Republic and Slovakia, and determine the extent of hybridization and introgression using nuclear (microsatellites and Randomly Amplified Polymorphic DNA, RAPD) and mitochondrial DNA (mtDNA) markers. Nuclear markers reveal hybrid zones between T. cristatus and T. dobrogicus at the foothills of the Carpathians in southern Slovakia, and between T. cristatus and T. carnifex in the southern parts of the Czech Republic. Analysis of mitochondrial cytochrome b sequences reveals T. cristatus and T. dobrogicus-specific haplotypes in contact zones in southern Slovakia. Surprisingly, most T. carnifex and individuals with mixed ancestry between T. carnifex and T. cristatus possess haplotypes specific for T. dobrogicus, most likely as a result of historical mtDNA introgression. Only one T. carnifex-specific haplotype carried by a single specimen is found in the Czech Republic. Our study shows that genetic structure of central European populations of crested newts is complex and influenced by historical and contemporary hybridization.
Overlapping measurements in the length of the genitalia of Leptidea sinapis/reali collected in Slovenia triggered an investigation of a possible natural hybridization between these two well known sibling species of butterflies. Random polymorphic DNA (RAPD) was used to generate species specific markers and sequences of the cytochrome oxidase subunit one gene for determination of the progeny. RAPD's clustering and mitochondrial DNA (mtDNA) phylogeny were congruent with the taxonomic placement of specimens of both species, but slightly incongruent with the results of the analysis of genital morphology. Two specimens with L. reali genitalia measurements, but genetically belonging to L. sinapis, had species specific RAPD markers of both species indicating probable hybrid origin. All the specimens with genitalia of intermediate length were also genetically assigned to L. sinapis indicating a possible one way introgression as predicted from their genitalia morphology. Leptidea sinapis was found predominantly in xerothermic habitats in Slovenia, whereas L. reali was more of a generalist except in the sub-Mediterranean region where it is limited to humid meadows.