Martina Podnar

Phylogeography of the Italian wall lizard, Podarcis sicula , as revealed by mitochondrial DNA sequences

In a phylogeographical survey of the Italian wall lizard, Podarcis sicula, DNA sequence variation along an 887‐bp segment of the cytochrome b gene was examined in 96 specimens from 86 localities covering the distribution range of the species. In addition, parts of the 12S rRNA and 16S rRNA genes from 12 selected specimens as representatives of more divergent cytochrome b haploclades were sequenced (together about 950 bp). Six phylogeographical main groups were found, three representing samples of the nominate subspecies Podarcis sicula sicula and closely related subspecies and the other three comprising Podarcis sicula campestris as well as all subspecies described from northern and eastern Adriatic islands. In southern Italy a population group with morphological characters of P. s. sicula but with the mitochondrial DNA features of P. s. campestris was detected indicating a probably recent hybridization zone. The present distribution patterns were interpreted as the consequence of natural events like retreats to glacial refuges and postglacial area expansions, but also as the results of multiple introductions by man.

Unusual Origin of a Nuclear Pseudogene in the Italian Wall Lizard: Intergenomic and Interspecific Transfer of a Large Section of the Mitochondrial Genome in the Genus Podarcis (Lacertidae)

Two distinct cytochrome b-like sequences were discovered in the genome of Podarcis sicula. One of them represents a nuclear copy of a mitochondrial sequence (numt-sic) differing by 14.3% from the authentic mitochondrial (mt) sequence obtained from the same individual. This numt, however, differs by only 2.7% from the mt sequence found in one population of Podarcis muralis, a related species in which no corresponding numt was detected. The numt-sic sequence extends over at least 7637 bp and is homologous to a section of the mt genome spanning from the tRNA-Lys to the tRNA-Pro gene. Premature mt stop codons were detected in two of the nine protein coding genes of numt-sic. The distribution of substitutions among the three codon positions and the transition/transversion ratio of the numt-sic sequence resemble, with few exceptions, those of functional mt genes, indicating a rather recent transfer to the nucleus. Phylogenetic analyses performed on the data set including P. sicula numt-cytb sequences as well as mt-cytb sequences from the same individuals and mt sequences of various P. muralis populations suggest that numt-sic originated in P. muralis. In a geographic survey, P. sicula populations belonging to different mt lineages, covering most of the distribution area, were screened for the presence of numt-sic and for a 15-bp duplication polymorphism in the numt-nd5 sequence. Our results suggest that numt-sic has spread rapidly through the species range via sexual transmission, thereby being transferred to populations belonging to well-separated mt lineages that diverged 1–3 Mya.

Two distinct evolutionary lineages of the Astacus leptodactylus species-complex (Decapoda : Astacidae) inferred by phylogenetic analyses

Abstract. Narrow-clawed crayfish (Astacus leptodactylus Eschscholtz, 1823 species-complex) is one of five European freshwater crayfish species. Even though widely distributed, it hasn’t been frequently studied and its taxonomy and systematics are unresolved. The results of a recent comparative morphometric character study revealed that morphometry of Asian and European populations differ significantly. In this research, for the first time, mitochondrial molecular markers (16S rRNA and COI) were used with the aim of elucidating the phylogenetic relationship between European and Asian populations of the narrow-clawed crayfish. Analyses included crayfish from Croatia, Bulgaria, Armenia, Russia, Poland and Turkey, and three different optimality criteria were applied. Phylogenetic relationships were reconstructed using the COI dataset, as well as the concatenated one (COI + 16S rRNA). For both datasets, congruent topologies were obtained and trees were characterised by the existence of two well supported phylogroups, one that included European populations, and the other Asian. Results indicate the presence of distinct evolutionary lineages within the A. leptodactylus species-complex, and corroborate previous results obtained using morphometric analyses.

Evolutionary history of relict Congeria (Bivalvia: Dreissenidae): unearthing the subterranean biodiversity of the Dinaric Karst

BackgroundPatterns of biodiversity in the subterranean realm are typically different from those encountered on the Earth’s surface. The Dinaric karst of Croatia, Slovenia and Bosnia and Herzegovina is a global hotspot of subterranean biodiversity. How this was achieved and why this is so remain largely unresolved despite a long tradition of research. To obtain insights into the colonisation of the Dinaric Karst and the effects of the subterranean realm on its inhabitants, we studied the tertiary relict Congeria, a unique cave-dwelling bivalve (Dreissenidae), using a combination of biogeographical, molecular, morphological, and paleontological information.ResultsPhylogenetic and molecular clock analyses using both nuclear and mitochondrial markers have shown that the surviving Congeria lineage has actually split into three distinct species, i.e., C. kusceri, C. jalzici sp. nov. and C. mulaomerovici sp. nov., by vicariant processes in the late Miocene and Pliocene. Despite millions of years of independent evolution, analyses have demonstrated a great deal of shell similarity between modern Congeria species, although slight differences in hinge plate structure have enabled the description of the two new species. Ancestral plesiomorphic shell forms seem to have been conserved during the processes of cave colonisation and subsequent lineage isolation. In contrast, shell morphology is divergent within one of the lineages, probably due to microhabitat differences.ConclusionsFollowing the turbulent evolution of the Dreissenidae during the Tertiary and major radiations in Lake Pannon, species of Congeria went extinct. One lineage survived, however, by adopting a unique life history strategy that suited it to the underground environment. In light of our new data, an alternative scenario for its colonisation of the karst is proposed. The extant Congeria comprises three sister species that, to date, have only been found to live in 15 caves in the Dinaric karst. Inter-specific morphological stasis and intra-specific ecophenotypic plasticity of the congerid shell demonstrate the contrasting ways in which evolution in the underground environments shapes its inhabitants.

DNA barcoding and first records of two rare Adicella species (Trichoptera: Leptoceridae) in Croatia

Abstract Two species of the genus Adicella, A. cremisa Malicky, 1972 and A. balcanica Botosaneanu & Novak,1965, were recorded in the summer period of 2014 which represent the first records of these species in Croatia. The former was collected at two relatively distant sites, the Krapina and the Zrmanja rivers, while the later was collected at the Krupa River. Both species were identified by morphological characteristics as well as DNA barcoding method. We calculated the uncorrected pairwise distances within Adicella and used molecular phylogenetic approach to delimit species. A. cremisa from Croatia showed no significant difference in the mtCOI region and they are highly similar to A. cremisa from Italy. Additionally, the ecological preferences and distribution of Adicella species are presented. Our findings represent a significant contribution to the aquatic biodiversity of the Western Balkans.

Molecular data do not confirm the Grecian anomalous blue Polyommatus (Agrodiaetus) aroaniensis (Brown, 1976) as a member of the Croatian fauna

The presence of the Grecian anomalous blue, Polyommatus (Agrodiaetus) aroaniensis (Brown, 1976) in Croatia has been recently published based on external morphology of collected specimens. The aim of this study was to evaluate the credibility of these findings by using DNA barcoding approach as the species exhibit phenotypic variability and can easily be misidentified with morphologically similar species Polyommatus (Agrodiaetus) ripartii. Therefore, we performed a molecular analysis of the mitochondrial gene for cytochrome c oxidase subunit I (COI) of the samples previously published as P. aroaniensis from different localities in Croatia. The results showed that all analysed samples belong to P. ripartii. Additionally, all analysed samples from different Croatian localities (Mt. Mala Kapela, Mt. Lička Plješivica, Mt. Poštak, Mt. Troglav and Mt. Kamešnica) together with previously published population of P. ripartii from Mt. Mosor belong to the same Eurasian clade of P. ripartii.

The first record of Tinodes antonioi Botosaneanu & Taticchi-Viganò, 1974 (Insecta, Trichoptera) in Croatia with DNA barcoding and ecological data and notice of biodiversity and distribution of the genus Tinodes in Croatia

1Department of Biology (Laboratory for Entomology), Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10 000 Zagreb, Croatia 2Croatian Natural History Museum, Demetrova 1, 10 000 Zagreb, Croatia 3Marina Tartaglie 2, 10 000, Zagreb, Croatia 4Grožnjanska 18, 10 000 Zagreb, Croatia 5Elektroprojekt, d.d., Civil and Architectural Engineering Department, Alexandera von Humboldta 4, 10 000 Zagreb, Croatia 6S. Toraigyrov State University in Pavlodar, Kazakhstan 7Tuk Bjelopoljski 14, Plitvička jezera, 53230, Korenica, Croatia

Preliminary data on genetics and morphometrics of Myotis alcathoe (Chiroptera, Vespertilionidae) in Croatia

Abstract Data on genetics and morphometrics of Myotis alcathoe from Croatia is presented.