Divergence Preceding Island Formation Among Aegean Insular Populations of the Freshwater Snail Genus Pseudorientalia (Caenogastropoda: Truncatelloidea) Author(s): Magdalena Szarowska, Sebastian Hofman, Artur Osikowski, and Andrzej Falniowski Source: Zoological Science, 31(10):680-686. 2014. Published By: Zoological Society of Japan DOI: http://dx.doi.org/10.2108/zs140070 URL: http://www.bioone.org/doi/full/10.2108/zs140070

BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.

BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.

ZOOLOGICAL SCIENCE 31: 680–686 (2014)

¤ 2014 Zoological Society of Japan

Divergence Preceding Island Formation Among Aegean Insular Populations of the Freshwater Snail Genus Pseudorientalia (Caenogastropoda: Truncatelloidea) Magdalena Szarowska1, Sebastian Hofman2, Artur Osikowski2, and Andrzej Falniowski1* 1

Department of Malacology, Institute of Zoology, Jagiellonian University, Gronostajowa 9, 30–387 Kraków, Poland 2 Department of Comparative Anatomy, Institute of Zoology, Jagiellonian University, Gronostajowa 9, 30–387 Kraków, Poland

Freshwater snails that inhabit islands are excellent model organisms for testing relationships between geological events and phylogeography, especially in the Aegean region. Although many Aegean islands were searched in the present study, species of the genus Pseudorientalia were only found on Lesvos, Samos, and Chios. Phylogenetic relationships between specimens living on these three islands were analysed using COI and 16S rRNA molecular markers and morphological data. A high level of diversity was found between islands. Genetic distances between clades showed differences high enough for the samples from different islands to be considered distinct species (p-distance: 0.105–0.133). These results are also supported by obvious morphological differences in shell morphology between islands. The mean divergence time between the Lesvos clade and Samos/Chios clade was 24.13 ± 3.30 Mya; between the Samos and Chios clades the divergence time was 14.80 ± 1.11 Mya. Our data suggest that high divergence may have occurred between Pseudorientalia populations during the Upper and Middle Miocene, when the Aegean region was part of a united landmass. It is possible that the observed highly divergent Pseudorientalia clades are relicts of high regional diversity that existed in the past. Key words:

Lesvos, Chios, Samos, Pseudorientalia, mtDNA, phylogeography, gastropods

INTRODUCTION The complex geological history of the Aegean area during the late Tertiary, especially the submergence and re-emergence of landmasses due to tectonic, volcanic, and eustatic events, significantly influenced the distribution of many terrestrial animals in this region (Poulakakis et al., 2005). During the Upper and Middle Miocene (23–12 Mya) the Aegean region was part of a united landmass (Dermitzakis and Papanikolaou, 1981). After the collision of the African and Arabian Plate with the Eurasian Plate at the end of the Middle Miocene, the formation of the Mid-Aegean trench began. This process ended during the Late Miocene (10–9 Mya) (Dermitzakis and Papanikolaou, 1981) and caused the separation of the Aegean islands into western and eastern groups. The Messinian salinity crisis (which started 6.5 Mya) was the second important geological event in the history of this area. After the closing up of the Strait of Gibraltar, the entire Mediterranean basin dried up, and the islands of this region became mountains in a steppe or desert, enabling overland migration between islands and mainland by terres* Corresponding author. Tel. : +48-12-664-5042; Fax : +48-12-664-5101; E-mail: [email protected] doi:10.2108/zs140070

trial animals. When the Strait of Gibraltar reopened (5.33 Mya), the basin was refilled from the Atlantic Ocean and most of the Mediterranean islands became isolated once more as they were surrounded by saltwater. With such a complex geological history in this region, the fate of particular islands is frequently complex and difficult to study (Poulakakis et al., 2005). Nevertheless, the analysis of fauna and flora inhabiting the Mediterranean islands gives a unique insight into processes of colonization and recolonization and their impact on the genetic structure of populations. Lesvos, Chios, and Samos belong to the eastern Aegean islands, and thus their fauna should be dominated by Asiatic organisms. The distance between Lesvos and Samos is 175 km, and Chios lies at about half this distance (Fig. 1). Moreover, connection between these islands and the landmass (Asia Minor) was maintained until the Middle Pleistocene (0.400–0.002 Mya) (Poulakakis et al., 2005). Low genetic divergence levels between animals occupying these islands may thus be expected. Terrestrial animals are excellent models for testing relationships between geological events and phylogeography, but freshwater snails have rarely been used as subjects in such studies. Radoman (1973) described a monotypic genus Pseudorientalia, belonging to his “Hydrobioidea”, now classified within the Hydrobiidae, subfamily Sadlerianinae

Pseudorientalia snails on Aegean Islands

681

Table 1. The sampling locations with their geographical coordinates and haplotypes for (Szarowska, 2006), and recorded superCOI and 16S rRNA mitochondrial genes detected in each locality. family Truncatelloidea, with its type species P. natolica (Küster, 1852) in Asia ID Site Coordinates COI haplotypes 16S haplotypes Minor (Radoman, 1983). Pseudorientalia Lesvos natolica is now known from a few localiN 39°21′41.1″ E 26°16′14.7″ L4×1, L5×1, L6×4 L2×2 L02 (G12L02) ties in northern Turkey, southeast of the Lesvos L1×4 N 39°21′30.5″ E 26°18′39.7″ L1×3, L2×1, L3×3 L01 Marmara Sea (Yildirim et al., 2006). (G12L01) Recently, P. tzekovi, a putative repreCH1×1, CH2×2, Chios CH1×7 N 38°33′28.7″ E 26°04′50.2″ CH01 sentative of the genus (since the female CH3×4 (G12CH01) reproductive organs were not examined) S1×1, S2×2, S3×1, Samos N 37°42′59.0″ E 26°46′51.3″ S02 S1×3 was described from northern Greece (G12S02) S4×1 (Glöer and Georgiev, 2012). We found another species of this genus, referred to as Pseudorientalia sp., on Samos Island (Szarowska et al., 2014). Unfortunately, there are neither DNA sequence data for P. natolica and P. tzekovi, nor are specimens available for morphological comparison with our material. Our extended fieldwork on the Aegean Islands, including Rhodes, Samos, Chios, Lesvos, Crete, Karpathos, Milos, Naxos, Paros, Mykonos, Tinos, Andros, Evvoia, and Kithira, also resulted in the discovery of Pseudorientalia on Chios and Lesvos. The objectives of this study were, first, to confirm, by analyses of morphological data, genus-level assignments of the four Aegean populations found and, second, to analyse the phylogenetic relationships between these populations by applying commonly used molecular markers (COI and 16S rRNA mitochondrial genes). The most interesting question was how these newly discovered populations differ morphologically and genetically. In the second part of the study we sought to estimate time of divergence between clades and to look for possible correlations of our phylogeFig. 1. Map of sampling sites. netic results with the geological history of the region. We focused on possible importance of major geological events, like formation of the Mid-Aegean trench, the Messinian salinity crisis and the Zanclean flood, which caused the isolation of Aegean islands. Finally, we propose and discuss most likely possible scenarios that could lead to current diversification of Pseudortientalia snails on Aegean Islands.

MATERIALS AND METHODS Biological material was collected from four populations across three islands (Fig. 1, Table 1): CH01, Manna Nerou spring, near Ghiosonas, Chios Island, 40 m a. s. l. (above sea level); L01, a spring near Klió, Epetimnos Mountain, Lesvos Island, 208 m a. s. l.; L02, a spring SSW of Argennos Epetimnos Mountain, Lesvos Island (Fig. 2), 362 m a. s. l.; S02, Despoti Fountain between Koutsi and Pirgos, Samos Island, 490 m a. s. l. Snails were washed twice and left to stand in 80% ethanol for around 12 hours. The ethanol was changed twice within 24 hours and finally, after a few days, the 80% solution was replaced with a 96% one. The samples were then stored at −20°C. The shells and soft parts were photographed with a CANON EOS 50D digital camera. For morphological comparisons five males and five females from each locality were dissected under a NIKON SMZ-U microscope. DNA was extracted from foot tissue. The tissue was hydrated in TE buffer (3 × 10 min.); total genomic DNA was then extracted using the SHERLOCK extracting kit (A&A Biotechnology), and the final product was dissolved in 20 μl TE buffer. The PCR reaction was performed with the following primers: LCOI490 (5′-GGTCAACAAATCATAAAGATATTGG-3′) (Folmer et al., 1994) and COR722b (5′-TAAACTTCAGGGTGACCAAAAAATYA-3′) (Wilke and Davis, 2000) for the cytochrome oxidase subunit I (COI) mito-

Fig. 2. tain.

Locality L02 – a spring SSW of Argennos Epetimnos Moun-

chondrial gene and LRJ12887 (5′-CCGGTCTGAACTCAGATCACGT-3′) and LRN13398 (5′-CGCCTGTTTAACAAAAACAT-3′) (Kasper et al., 2004) for the 16S mitochondrial ribosomal gene. The PCR conditions were as follows: COI: initial denaturation step of 4 min at 94°C, followed by 35 cycles of 1 min at 94°C, 1 min at 55°C, 2 min at 72°C, and a final extension of 4 min at 72°C; 16S: initial denaturation step of 2 min 30 s at 90°C, followed by 10 cycles of 50 s at 92°C, 50 s at 43°C, and 40 s at 72°C, and 36 cycles of

682

M. Szarowska et al.

30 s at 92°C, 40 s at 44.4°C, 40 s at 72°C. The total volume of each PCR reaction mixture was 50 μl. To check the quality of the PCR products, 10 μl was run on 1% agarose gel. The PCR products were purified using Clean-Up columns (A&A Biotechnology) and were then amplified in both directions (Hillis et al., 1996) using BigDye Terminator v3.1 (Applied Biosystems). The sequencing reaction products were purified using ExTerminator Columns (A&A Biotechnology); DNA fragments then underwent electrophoresis on an ABI Prism sequencer. Sequences were initially aligned in the ClustalW program in MEGA 6 (Tamura et al., 2013) and then checked manually in Bioedit 7.1.3.0 (Hall, 1999). Basic sequence statistics including haplotype polymorphism and nucleotide divergence were calculated in DnaSP 5.10 (Librado and Rozas, 2009). The mutational saturation, examined using DAMBE (Xia, 2013) by saturation test by Xia et al. (2003) and by plotting the numbers of transitions and transversions against the percentage sequence divergence, revealed no saturation in any of sequences studied. We performed phylogeny reconstruction for COI and 16S rRNA separately, using Daphniola exigua Fig. 3. Shells of Pseudorientalia. (A–C) Chios Island (locality CH01); (D–J) (Schmidt, 1856) (JF916467, Falniowski and Szarowska, Lesvos Island (D, F, G, I): locality L01; (E, H, J) locality L02); (K–N) Samos 2011a) and Sadleriana fluminensis (Küster, 1852) Island (locality S02). Scale bar: 1 mm. (AY222657, Szarowska and Wilke, 2004) as outgroups, respectively. For COI gene relationship analysis, the sequence of Tefennia tefennica Schütt and Yildirim, 2003 from Turkey (JX982804, Ça˘glan et al., 2012) was also used. Best-fit models of nucleotide evolution for COI genes were selected following the AICc criteria in jModelTest v. 2.1.4 (Guindon and Gascuel, 2003; Darriba et al., 2012). The maximum likelihood approach was used to construct a phylogenetic tree with PhyML 3.0 (Guindon and Gascuel, 2003). To infer haplotype networks a median-joining calculation was implemented in NETWORK 4.6.1.1 (Bandelt et al., 1999). To test the molecular clock, the combined data from COI and 16S rRNA were used. To check whether these two datasets could be analyzed together, a partition homogeneity test (Farris et al., 1995) was performed with PAUP*4.0 (Swofford, 2002). This test did not reveal significant differences (P = 1.000; 10,000 replicates). Two hydrobiids, Peringia ulvae Pennant, 1777 and Salenthydrobia ferreri Wilke, 2003 (AF478401, AF478410; Wilke, 2003) were used as outgroups. The divergence time between these two species was used to calibrate the molecular clock, with correction according to Falniowski et al. (2008). The likelihood for trees with and without the molecular clock assumption for a Likelihood Ratio Test (LRT) (Nei and Kumar, 2000; Posada, 2003) were calculated with PAUP. The Relative Rate Test (RRT) (Tajima, 1993) was performed in MEGA. Time estimates were calculated using a non-parametric rate smoothFig. 4. Soft part morphology and anatomy. (A) head of the female ing (NPRS) analysis with the recommended Powell algorithm, as from Lesvos Island (locality L01); (B) penis of specimen from Chios implemented in the software r8s (v.1.7 for Linux) (Sanderson, 1997, Island; (C) female reproductive organs from the same locality. Scale 2003). bars: 0.5 mm.

RESULTS The shells of our Pseudorientalia (Fig. 3), which resemble those photographed and described for P. natolica, differ slightly between the two populations from Lesvos (Fig. 3A– G: compare Fig. 3A, C, D, F from L01 vs Fig. 3B, E, G from L02). All, however, differ from those on the islands of Chios (Fig. 3H–J) and Samos (Fig. 3K–N). In Samos, some specimens were larger and relatively high-spired (Fig. 3K). Figure 4A shows the head, on which there are large eyes with dark spots of black pigment around them, and black pigment proximally at the proboscis. The penis (Figs. 4B,

5E–F) is simple, without outgrowths, and bent, with a rather long tip and black pigment at its medial part. The distal part of the female reproductive organs (Figs. 4C, 5A–D) shows no differences between the studied populations, and is characterized by two seminal receptacles, one of which (rs1 according to Radoman, 1983) is small or nearly vestigial. The oviduct shows a rather prominent loop, and there is a bulky, nearly spherical bursa copulatrix (sometimes approaching the size of the accessory duct complex). The 25 specimens from the four populations sequenced

Pseudorientalia snails on Aegean Islands

683

for COI (552 bp, GenBank Accession numbers: KJ920473– KJ920497) were grouped into 14 haplotypes arranged in three main mitochondrial clades, each of which occupied one of the study islands (Fig. 6, Table 1). The AIC selected the model TrN+G, with base frequencies of A = 0.2697, T = 0.3963 C = 0.1609, G = 0.1731, and gamma parameter = 0.1440. The phylogenetic analysis of COI indicated a high level of diversity between the populations, but low diversity within them (Fig. 6). The two populations from Lesvos are more distant from the other two, and the haplotype number is the highest within this group. The populations from Samos and Chios are more closely related to one another, but Tefennia tefennica clusters between them. The high diversity level between islands is also confirmed by the 16S rRNA gene (312 bp, GenBank Accession numbers: KJ920457–KJ920472) analysis (Fig. 7). The AIC selected the model HKY+G, with base frequencies of A = 0.259, T = 0.389 C = 0.168, G = 0.183, and gamma parameter = 0.17. From 16 studied specimens, only four haplotypes were found, each characteristic of a single population (Table 1). Genetic distances between the three clades for COI and 16S rRNA (Table 2) are high enough to be characteristic of distinct species (0.105–0.133). For combined COI and 16S rRNA sequences, the AIC

Fig. 5. Female and male reproductive organs of Pseudorientalia. (A–D) distal part of female reproductive organs; (A, B) specimens from Lesvos Island (A) locality L01, (B) locality L02; (C, D) the same specimen from Chios Island (locality CH01); (E, F) penes of specimens from Lesvos Island (locality L02). Scale bar: 0.5 mm.

Fig. 7. The maximum–likelihood phylogram (HKY + G, 10,000 bootstrap replicates) for 16S rRNA gene. The ML tree was rooted by using the Sadleriana fluminensis.

Fig. 6. The maximum–likelihood phylogram (TrN+G, 10,000 bootstrap replicates) for COI gene. The ML tree was rooted by using the Daphniola exigua. The median–joining haplotype networks were shown, for each clade occupied one of the study islands.

684

M. Szarowska et al.

Table 2. P-distance values for COI (below diagonal) and 16S rRNA (above diagonal).

S02 CH01 L01 L02

S02

CH01

L01

L02

– 0.077 0.133 0.133

0.007 – 0.120 0.118

0.108 0.108 – 0.005

0.105 0.105 0.003 –

with jModelTest found model HKY + G, base frequencies: A = 0.3110, T = 0.3764 C = 0.1487, G = 0.1640 and gamma parameter = 0.093. Tajima’s RRTs for each pair of Pseudorientalia haplotype with either Salenthydrobia or Peringia as outgroup did not reject the molecular clock hypothesis. However, the LRT test (LRT = 101.39, df = 10, P < 0.0001) rejected the equal evolutionary rate throughout the tree for this dataset. For this reason, non–parametric rate smoothing (NPRS) analysis with the recommended Powell algorithm was used. Because isolation of the Atlantic Peringia from the Mediterranean Salenthydrobia must have begun when the Mediterranean Basin started to separate from the Atlantic (Falniowski et al., 2008), the calibration point was established at 5.96 Mya. Applying this calibration, the mean divergence time between the Lesvos clade and the Samos/Chios clade was 24.13 ± 3.30 Mya. Between the Samos and Chios clades this time was 14.80 ± 1.11 Mya. DISCUSSION The shells examined in this study resemble the one photographed by Radoman (1983) for P. natolica, although they were generally smaller than those seen in the figures in this previous study. Within the Trunatelloidea, the shell is a poor taxonomic character, being simple and highly variable. However, differences visible between the shells of the snails from the three islands in the present study suggest that each island is inhabited by a distinct species. The penis was similar to that already described and illustrated for the population from Samos (Szarowska et al., 2014), and to the drawings of Radoman (1983) and photograph provided by Glöer and Georgiev (2012). In fact, such a simple penis is hardly diagnostic, but its morphology confirms that all the Aegean populations belong to the genus Pseudorientalia. Similar remarks can be made about the female reproductive organs: in all the studied populations they present the same characteristics, shared with those described for the Samos populations (Szarowska et al., 2014), and those of P. natolica given by Radoman (1983). In fact, similar female reproductive organs can also be found in Turcorientalia Radoman, 1973, but the penis, with a characteristic double outgrowth in the latter genus (Radoman, 1983), is different from the one found in our Aegean populations. The clustering of Tefennia tefennica within the clade consisting of our Pseudorientalia from Chios and Samos in the molecular COI tree is somewhat unexpected. Considering the morphology, the penis of T. tefennica has a small outgrowth, and there is no distal receptaculum seminis (rs1) (Ça˘glan et al., 2012). A non-prominent penile outgrowth may or may not be present in the same species: Horatia klecakiana (Bourguignat, 1887) may be a good example (Radoman, 1983). In contrast, the presence of receptacula was unexpectedly shown to be very

stable throughout the phylogeny within the Truncatelloidea (Szarowska, 2006). However, in our Pseudorientalia, the rs1 are small, nearly vestigial, and it is also possible that such a vestigial rs1 could have been overlooked in Tefennia. In any case, all the morphological data confirm that all the Aegean populations belong to the same genus, Pseudorientalia. Pseudorientalia snails were only found on the three islands of the east Aegean area described in this paper. Despite an extensive search, no members of this genus were found on the islands to the west of the Mid-Aegean trench. This may indicate that the Lesvos, Chios, and Samos populations of Pseudorientalia are the relict of a land connection of these islands with Asia Minor. Since most Pseudorientalia populations are found in Turkey (Yildirim et al., 2006), it is possible that Pseudorientalia only inhabits areas to the east of the Mid-Aegean trench and are separated from western Aegean islands. This is not an uncommon pattern, since the isolation between western and eastern Aegean islands, due to the formation of the Mid-Aegean trench, has been reported for many animals: e.g. land snails (Zonites: Kornilios et al., 2009; Mastus: Parmakelis et al., 2005), scorpions (Parmakelis et al., 2006a), Coleoptera (Fattorini, 2002), and reptiles (Lymberakis and Poulakakis, 2010). According to the p-distance values for COI, each island harbours a distinct species (e.g. Falniowski and Szarowska, 2011a). High divergence levels between populations inhabiting studied islands may be surprising, since the connection between these islands probably disappeared very recently, in the Middle Pleistocene. Estimates of the time of divergence between Lesvos and Chios/Samos populations located this event before the formation of the Mid-Aegean trench. However, with one-point calibration, molecular clock estimates are prone to significant errors, and reasonable estimates of confidence intervals are unavailable. On the other hand, this point of calibration was used several times (e.g., Falniowski et al., 2008; Smole´n and Falniowski, 2009; Falniowski and Szarowska, 2011a) producing results consistent with geological history. The previously described snail Tefennia tefennica from Turkey (Ça˘g lan et al., 2012) belongs to the same clade as Pseudorientalia from Chios and Samos; snails from Lesvos formed a separate clade. These two facts may indicate that the snails from Lesvos belong to another genus, but our morphological data do not support this idea. The population from Lesvos also belongs to Pseudorientalia, but is more divergent from the Chios and Samos populations. It is possible that two highly divergent Pseudorientalia clades exist, one from continental Greece, and the second from the east Aegean Islands and Minor Asia, or—considering geography alone—that the species from continental Greece is more related to the one from Lesvos. The population from Lesvos could have originated from the western part of the Aegean Sea, and then migrated eastward in some way. Unfortunately, extremely little is known about the biology of the Pseudorientalia. For other snails, a few mechanisms of island colonization have been described, such as bird transportation (Gittenberger et al., 2006; Wada et al., 2012), or introduction by man. This last mechanism was described, for example, for the Lesvos population of the land snail Zonites (Kornilios et al., 2009),

Pseudorientalia snails on Aegean Islands

although this is hardly probable in case of a stenothermic species inhabiting springs. Bird transportation seems very probable in the Greek Bythinella (Falniowski and Szarowska, 2011b), which is quite resistant to desiccation (Falniowski, 1987; Szarowska, 2000), whereas the majority of spring malacofauna are not. It should be noted, that the Pseudorientalia described in this paper were found on very few Aegean islands, which strongly suggests that the migration potential of these snails is low, although the factors (whether ecological, morphological or physiological) responsible for this phenomenon remain unknown. For this reason, strong limitation of gene flow between island populations may be an important factor maintaining significant genetic divergence between Pseudorientalia inhabiting Samos, Chios and Lesvos. It is possible that the observed Pseudorientalia clades are relicts of a high regional diversity (maybe from the Miocene), since estimated time of divergence between Samos and Chios clades also indicates an old separation between these two clades, close to the ending of forming of the Mid-Aegean trench. In the eastern part of the Aegean area there could have existed three or more (e.g., from Asia Minor) clades, and some clades could have existed in continental Greece. Unfortunately there are no molecular data for a putative Pseudorientalia species from continental, northern Greece (Glöer and Georgiev, 2012). In our study, shell morphology and DNA sequence data strongly suggest that the three island populations are distinct species. If this is true, we can hypothesize that gene pools of these “species” could have been at least partly isolated long before the connection between islands disappeared. In such a case, the current highly divergent clades could be relicts of high ancestral polymorphism of Pseudorientalia snails. Our data suggest that high divergence between Pseudorientalia populations may have occurred during the Upper and Middle Miocene, when the Aegean region was part of a united landmass. The formation of the Mid-Aegean trench may also have affected distribution and diversification of these snails, since almost all recent Pseudorientalia populations occur on east Aegean islands and in Asia Minor. This old divergence of the Pseudorientalia was, of course, reinforced by divergence after isolation of islands by seawater, after the Zanclean flood. For snails living in springs, exposed on severe bottlenecks, or subject to diversified selection, etc., such allopatric diversification is the strong factor increasing the divergence level. When using geological age of islands to calibrate molecular clock, it is important to remember that some taxa may be older than the islands they inhabit. It may be a consequence of species’ survival on former nearby islands that have been submerged or on a mainland (Heads, 2011). Such a situation is not unlikely in the case of Pseudorientalia, because Lesvos, Samos, and Chios are near the mainland and surrounded by other, smaller islands. Such a phenomenon may explain the presence of such divergent populations of Pseudorientalia on relatively closely situated islands. High regional diversities of land snails in the Mediterranean region have often been reported (Bank et al., 1998; Cameron et al., 2000). They are substantially higher in comparison with the northern part of Europe, or at the same latitudes in North America (Cameron et al., 2000). This phenom-

685

enon may be a relict of Miocene differentiations, also before the opening of the Mid-Aegean Trench (e.g., Kasapidis et al., 2005; Steinfartz et al., 2000; Parmakelis et al., 2006b). Some of this differentiation can be attributed to the long– term fragmentation of landmasses caused by tectonics. Some other factors could also have made a substantial contribution to this diversity, like the greater range of habitable environments available in the Aegean region, microgeographical differentiation within single habitat types and climatic fluctuations (e.g., in humidity and temperature). ACKNOWLEDGMENTS We thank two anonymous reviewers for useful comments. This study was supported by a grant from the National Science Centre [2011/01/B/NZ8/01721] to Andrzej Falniowski.

REFERENCES Bandelt HJ, Forster P, Röhl A (1999) Median-joining networks for inferring intraspecific phylogenies. Mol Biol Evol 16: 37–48 Bank RA, Falkner G, Gittenberger E, Hausdorf B, Proschwitz T, Ripken TEJ (1998) Biodiversity of the western Palearctic region as exemplified by continental Mollusca. In: “Abstracts, World Congress of Malacology, Washington, DC” Ed by R Bieler, PM Mikkelsen, 25, Unitas Malacologica, Chicago Bourguignat JR (1887) Étude sur les noms génériques des petites paludinidées a opercule spirescent suivie de la description du nouveau genre Horatia. V. Tremblay, Paris Ça˘glan DC, Yildirim MZ, Szarowska M, Falniowski A (2012) Phylogenetic position of Tefennia Schütt et Yildrim, 2003 (Casenogastropoda: Rissooidea). Folia Malacol 20: 271–277 Cameron RAD, Mylonas M, Vardinoyannis K (2000) Local and regional diversity in some Aegean land snail faunas. J Moll Stud 66: 131–142 Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Meth 9: 772 Dermitzakis MD, Papanikolaou DJ (1981) Palaeogeography and geodynamics of the Aegean region during the Neogene. Annales Géologique des Pays Hellénique 30: 245–289 Falniowski A (1987) Hydrobioidea of Poland (Prosobranchia: Gastropoda). Folia Malacol 1: 1–122 Falniowski A, Szarowska M (2011a) The genus Daphniola Radoman, 1973 (Caenogastropoda: Hydrobiidae) in the Peloponnese, Greece. Folia Malacol 19: 131–137 Falniowski A, Szarowska M (2011b) Radiation and phylogeography in a spring snail Bythinella (Mollusca: Gastropoda: Rissooidea) in continental Greece. Ann Zool Fenn 48: 67–90 Falniowski A, Szarowska M, Sirbu I, Hillebrand A, Baciu M (2008) Heleobia dobrogica (Grossu & Negrea, 1989) (Gastropoda: Rissooidea: Cochliopidae) and the estimated time of its isolation in a continental analogue of hydrothermal vents. Moll Res 28: 165–170 Farris JS, Källersjö M, Kluge AG, Bult C (1995) Testing significance of incongruence. Cladistics 10: 315–319 Fattorini S (2002) Biogeography of the tenebrionid beetles (Coleoptera: Tenebrionidae) on the Aegean Islands (Greece). J Biogeogr 29: 49–67 Folmer O, Black M, Hoeh W, Lutz RA, Vrijenhoek RC (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotech 3: 294–299 Gittenberger ED, Groenenberg SJ, Kokshoorn B, Preece RC (2006) Molecular trails from hitch-hiking snails. Nature 439: 409 Glöer P, Georgiev D (2012) Three new gastropod species from Greece and turkey (Mollusca: Gastropoda: Rissooidea) with

686

M. Szarowska et al.

notes on the anatomy of Bythinella charpentieri cabirius Reischütz 1988. North–West J Zool 8: 278–282 Guindon S, Gascuel O (2003) A simple, fast and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52: 696–704 Hall TA (1999) BioEdit: a user–friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids SympSer 41: 95–98 Heads M (2011) Old taxa on young islands: a critique of the use of island age to data island-endemic clades and calibrate phylogenies. Syst Biol 60: 204–218 Hillis DM, Mable BK, Larson A, Davis SK, Zimmer EA (1996) Nucleic acids IV: sequencing and cloning. In “MolecularSystematics. 2 ed.” Ed by DM Hillis, C Moritz, BK Mable, Sinauer Associates, Inc., Sunderland, Massachusetts, pp. 321–381 Kasapidis P, Magoulas A, Mylonas M, Zouros E (2005) The phylogeography of the gecko Cyrtopodionkotschyi (Reptilia: Gekkonidae) in the Aegean archipelago. Mol Phylogenet Evol 35: 612–623 Kasper ML, Reeson AF, Cooper SJB, Perry KD, Austin AD (2004) Assessment of prey overlap between a native (Polistes humilis) and an introduced (Vespula germanica) social wasp using morphology and phylogenetic analyses of 16S rDNA. Mol Ecol 13: 2037–2048 Kornilios P, Poulakakis N, Mylonas M, Vardinoyannis K (2009) The phylogeny and biogeography of the genus Zonites Montfort, 1810 (Gastropoda: Pulmonata): preliminary evidence from mitochondrial data. J Moll St 75: 109–117 Küster HC (1852) Die Gattungen Paludina, Hydrocaena und Valvata. In “Systematische Conchologie—Cabinet von Martini und Chemnitz 1(21)” Ed by GH Schubert, JA Wagner, Bauer und Raspe, Nürnberg, pp 1–95 Librado P, Rozas J (2009) DnaSP v5 a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25: 1451– 1452 Lymberakis P, Poulakakis N (2010) Three continents claiming an Archipelago: The evolution of Aegean’s herpetofaunal diversity. Diversity 2: 233–255 Nei M, Kumar S (2000) Molecular Evolution and Phylogenetics. Oxford University press, Oxford, UK and New York Parmakelis A, Pfenninger M, Spanos L, Louis C, Mylonas M (2005) Inference of radiation in Mastus (Gastropoda, Pulmonata, Enidae) on the island of Crete. Evolution 59: 991–1005 Parmakelis A, Stathi I, Spanos L, Papagiannakis G, Louis C, Mylonas M (2006a) Phylogeography of Iurus dufoureius (Brulle, 1832) (Scorpiones, Iuridae). J Biogeogr 33: 251–260 Parmakelis A, Stathi I, Chatzaki M, Simaiakis S, Spanos L, Louis C, Mylonas M (2006b) Evolution of Mesobuthus gibbosus (Brulle, 1832) (Scorpiones: Buthidae) in the northeastern Mediterranean region. Mol Ecol 15: 2883–2894 Posada D (2003) Selecting models of evolution. In “The Phylogenetic Handbook. A Practical Approach to DNA and Protein Phylogeny” Ed by M Salemi, AM Vandamme, Cambridge University Press, Cambridge, UK, pp. 256–282 Poulakakis N, Lymberakis P, Tsigenopoulos CS, Magoulas A, Mylonas M (2005) Phylogenetic relationships and evolutionary history of snake-eyed skink Ablepharus kitaibelii (Souria: Scincidae). Mol Phylogenet Evol 34: 245–256 Radoman P (1973) New classification of fresh and brackish water Prosobranchia from the Balkans and Asia minor. Posebna

Izdanja, Prirodn Mus Beograd 32: 1–30 Radoman P (1983) Hydrobioidea a superfamily of Prosobranchia (Gastropoda). I Systematics. Monographs 547/57. Department of Sciences Serbian Academy of Sciences and Arts Beograd Sanderson MJ (1997) A nonparametric approach to estimating divergence times in the absence of rate constancy. Mol Biol Evol 14: 1218–1231 Sanderson MJ (2003) R8s: inferring absolute rates of molecular evolution, divergence times in the absence of a molecular clock. Bioinformatics 19: 301–302 Smole´n M, Falniowski A (2009) Molecular phylogeny and estimated time of divergence in the Central European Melanopsidae: Melanopsis, Fagotia and Holandriana (Mollusca: Gastropoda: Cerithioidea). Folia Malacol 17: 9–11 Steinfartz S, Veith M, Tautz D (2000) Mitochondrial sequence analysis of Salamandra taxa suggests old splits of major lineages and postglacial recolonizations of Central Europe from distinct source populations of Salamandra salamandra. Mol Ecol 9: 397–410 Swofford DL (2002) PAUP* – Phylogenetic analysis using parsimony (*and other methods). Ver. 4. [Computer software and manual]. Sinauer Associates Inc., Sunderland, Massachusetts Szarowska M (2000) Environmental threats and stability of Bythinella populations in SouthPoland (Gastropoda: Prosobranchia: Hydrobioidea). Malakol Ab Staat Mus Tier Dresden 20: 93–98 Szarowska M (2006) Molecular phylogeny, systematics and morphological character evolution in the Balkan Rissooidea (Caenogastropoda). Folia Malacol 14: 99–168 Szarowska M, Wilke T (2004) Sadleriana pannonica (Frauenfeld, 1865): a lithoglyphid, hydrobiid or amnicolid taxon? J Mollusc Dtud 70: 49–57 Szarowska M, Hofman S, Osikowski A, Falniowski A (2014) Pseudorientalia Radoman, 1973 (Caenogastropoda: Rissooidea) on Samos Island, Aegean Sea. Folia Malacol 22: 11–20 Tajima F (1993) Simple methods for testing molecular clock hypothesis. Genetics 135: 599–607 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30: 2725–2729 Wada S, Kawakami K, Chiba S (2012) Snails can survive passage through a bird’s digestive system. J Biogeogr 39: 69–73 Wilke T (2003) Salenthydrobia gen. nov. (Rissooidea: Hydrobiidae): a potential relict of the Messinian sality crisis. Zool J Linn Soc 137: 319–336 Wilke T, Davis GM (2000) Infraspecific mitochondrial sequence diversity in Hydrobia ulvaeand Hydrobia ventrosa (Hydrobiidae: Rissoacea: Gastropoda): Do their different life histories affect biogeographic patterns and gene flow? Biol J Linn Soc 70: 89– 105 Xia X (2013) DAMBE5: A comprehensive software package for data analysis in molecular biology and evolution. Mol Biol Evol 30: 1720–1728 Xia X, Xie Z, Salemi M, Chen L, Wang Y (2003) An index of substitution saturation and its application. Mol Phylogenet Evol 26: 1–7 Yildirim MZ, Koca SB, Kebapçi Ü (2006) Supplement to the Prosobranchia (Mollusca: Gastropoda). Fauna of fresh and brackish waters of Turkey. Turk J Zool 30: 197–204 (Received April 3, 2014 / Accepted June 19, 2014)

Divergence preceding island formation among Aegean insular populations of the freshwater snail genus Pseudorientalia (Caenogastropoda: Truncatelloidea).

Freshwater snails that inhabit islands are excellent model organisms for testing relationships between geological events and phylogeography, especiall...
317KB Sizes 1 Downloads 6 Views