Daniela Guicking

A new cryptic species of pond turtle from southern Italy, the hottest spot in the range of the genus Emys (Reptilia, Testudines, Emydidae)

Geographic variation in the mtDNA haplotypes (cytochrome b gene) of 127 European pond turtles from Italy was investigated. Thirty‐eight of the Italian samples were also studied by nuclear fingerprinting (ISSR PCR) and compared with samples from other parts of the range representing all nine currently known mtDNA lineages of Emys orbicularis. Our genetic findings were compared against morphological data sets (measurements, colour pattern) for 109 adult turtles from southern Italy. Italy is displaying on a small geographical scale the most complicated variation known over the entire distributional area of Emys (North Africa over Europe and Asia Minor to the Caspian and Aral Seas). The Tyrrhenic coast of the Apennine Peninsula, the Mt. Pollino area and Basilicata are inhabited by Emys orbicularis galloitalica, a subspecies harbouring a distinct mtDNA lineage. The same lineage is also found in Sardinia. Along the Adriatic coast of Italy and on the Salentine Peninsula (Apulia, southern Italy), another morphologically distinctive subspecies (Emys orbicularis hellenica) occurs, which also bears a different mtDNA lineage. A higher diversity of mtDNA haplotypes in the south of the Apennine Peninsula suggests that the glacial refugia of E. o. galloitalica and E. o. hellenica were located here. A further refuge of E. o. hellenica probably existed in the southern Balkans. The west coasts of the Balkans and Corfu have probably been colonized from Italy and not from the geographically closer southern Balkanic refuge. In Sicily, a third mtDNA lineage is distributed, which is sister to all other known lineages of Emys. Morphologically, Sicilian pond turtles resemble E. o. galloitalica. However, nuclear fingerprinting revealed a clear distinctiveness of the Sicilian taxon, whereas no significant divergence was detected between representatives of the other eight mtDNA lineages of Emys. Furthermore, nuclear fingerprinting provided no evidence for current or past gene flow between the Sicilian taxon and the mainland subspecies of E. orbicularis. Therefore, Sicilian pond turtles are described here as a species new to science. Some populations in Calabria and on the Salentine Peninsula comprise individuals of different mtDNA lineages. We interpret this as a natural contact. However, we cannot exclude that these syntopic occurrences are the result of human activity. For example, in other parts of Italy, the natural distribution pattern of Emys is obscured by allochthonous turtles. This could also be true for southern Italy. The discovery of the complex taxonomic differentiation in southern Italy requires reconsidering conservation strategies.

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.

Comparative chloroplast DNA phylogeography of two tropical pioneer trees, Macaranga gigantea and Macaranga pearsonii (Euphorbiaceae)

Macaranga (Euphorbiaceae) has received much ecological and evolutionary research attention as a genus that includes some of the most conspicuous pioneer trees of Southeast Asian tropical rainforests and because of its manifold associations with ants, including about 30 species that are obligate ant-plants (myrmecophytes). We used sequence data from three chloroplast DNA loci (ccmp5, ccmp6, atpB-rbcL) to assess phylogeographical patterns in species of Macaranga, section Pruinosae, sampled from various regions of Borneo and the Malay Peninsula. Forty-nine chloroplast DNA haplotypes (HT) were identified among 768 specimens from five species, Macaranga gigantea (N = 329; 23 HT), Macaranga pearsonii (N = 347; 21 HT), Macaranga puberula (N = 24; 4 HT), Macaranga hosei (N = 48; 6 HT), and Macaranga pruinosa (N = 20; 5 HT). Forty-one haplotypes were species-specific, whereas eight haplotypes were shared by two, three, or four species and occupied internal positions in a parsimony network. Population genetic parameters based on haplotype frequencies proved to be in a similar range in the non-myrmecophytic M. gigantea and in the ant-associated M. pearsonii, which have overlapping distributions in northern and eastern Borneo. A comparison of GST and NST values revealed a strong phylogeographic structure in both species, whereas colonization pathways suggested by the network topology were different. Both species exhibited similar levels of haplotypic diversity and moderate to high levels of population differentiation. There were no obvious indications for an influence of the symbiotic ant partners on the population structure of their host plants.

Morphometrics of the pink-footed shearwater (Puffinus creatopus): influence of sex and breeding site

We present morphometric data for the pink-footed shearwater (Puffinus creatopus) from both Chilean breeding colonies on Isla Mocha and Juan Fernández: weight, total length, wing length, tail length, head length, three bill and three tarsus size measurements and length of middle toe. Significant differences were found for most measurements between males and females as well as between birds from the two different breeding localities. In general, males were bigger than females, and birds from Juan Fernández were bigger than those from Isla Mocha. Geographic differences most likely reflect some kind of ecomorphological adaptation. Some variables, especially the total length of head, seem to be useful for sex determination in the field.

Satellite tracking of the Pink-footed Shearwater in Chile

-Satellite tracking was used to examine movements of the threatened Pink-footed Shearwater (Puffinus creatopus) from Isla Mocha, Chile, during the chick-rearing period in 1998. We were able to track one breeding bird, one bird of unknown status and one prospector. Whereas the first two performed foraging flights and returned to the colony after about a two weeks absence, the last traveled north for four weeks, suggesting that it had started migration towards the wintering grounds in the northern hemisphere. Faster travel speeds during the day than during the night were found for all three birds. A major foraging zone was located about 250-300 km north of Isla Mocha and close to the Chilean mainland. The preference of Pink-footed Shearwaters for waters close to the coast when feeding, as well as when migrating, has implications for the conservation of the species. Received 5July 2000, accepted 26 August 2000.

Genetic variability in wild populations of Prunus divaricata Ledeb. in northern Iran evaluated by EST-SSR and genomic SSR marker analysis

A population genetic analysis based on eight genomic SSR markers and three EST-SSR (expressed sequence tags) markers developed in peach (Prunus persica (L.) Batsch) and Japanese plum (Prunus salicina Lindl.) was carried out in 12 wild populations of cherry plum (Prunus divaricata Ledeb.) sampled along the Iranian coast of the Caspian Sea. A total of 184 alleles (3–31 per locus) were detected with a mean value of 16.7 alleles per locus. None of the loci or populations showed deviation from Hardy–Weinberg equilibrium, and all markers proved to be unlinked. The mean values for the observed and the expected heterozygosity were 0.66 and 0.73, respectively. There was very little genetic differentiation among populations, as was indicated by low overall values of Wright’s FST (0.03) and Nei’s GST (0.08). An analysis of molecular variance (AMOVA) showed that 96.8% of the total variance was attributable to differences between individuals within populations. Genetic and geographic distances were nevertheless positively correlated, as evidenced by a Mantel test. The high level of genetic diversity and the apparent lack of genetic structure in wild P. divaricata may be attributed to frequent long distance gene flow through frugivorous birds and possibly humans, as has been documented for other Prunus species.

Relict Populations and Endemic Clades in Palearctic Reptiles: Evolutionary History and Implications for Conservation

The phylogeographic history of eight species complexes of West Palearctic reptiles was reconstructed using mitochondrial and nuclear markers. Cryptic endemic taxa were detected in the Southern European peninsulas (Iberia, Southern Italy/Sicily, and Greece) as well as in North Africa, Anatolia, Iran, and the Caucasus. These endemics are mainly of Tertiary or early Pleistocene age. Only part of them can be categorized as relicts of a former, more widely distributed taxon, having survived in ice age refugia; others have probably always remained in a restricted area, close to their origin.

Single nucleotide sequence analysis: a cost‐ and time‐effective protocol for the analysis of microsatellite‐ and indel‐rich chloroplast DNA regions

We present a simple method to screen for DNA sequence variation in microsatellite‐ and indel‐rich regions of the chloroplast genome. The single nucleotide sequence (SNS) analysis provides a trade‐off between the time‐ and cost‐effective, but less informative and homoplasy‐sensitive electrophoretic detection of microsatellite and indel size variation on the one hand, and more costly, but also more accurate methods like DNA sequencing on the other. The principle of the SNS method is to sequence one instead of all four nucleotides of a target region amplified by polymerase chain reaction. By careful selection of the respective nucleotide, almost the same amount of information can be retrieved from these partial sequences as could be from complete sequences; however, only a third to a fourth of the money and time resources are needed.

Mechanisms of Speciation in Southeast Asian Ant-Plants of the Genus Macaranga (Euphorbiaceae)

The palaeotropic genus Macaranga (Euphorbiaceae) is an excellent model system to analyze co-evolutionary processes associated with myrmecophytism, a mutualistic interaction between plants and ants. Ant-plants like Macaranga provide nesting space and feed their partners, whereas the ants protect the plants from herbivores and competitors such as lianas. We used genome-based evidence to investigate speciation mechanisms in Macaranga ant-plants, and their co-evolution with ants from the genus Crematogaster. Our previous work had shown that myrmecophytic Macaranga species show little genetic differentiation, suggesting an adaptive radiation. We hypothesized that the obligatory symbiosis with ants may reduce gene flow among plant populations, eventually enhancing allopatric speciation. To test this hypothesis, we verified the monophyly of the investigated plant lineages by phylogenetic analyses, reconstructed parsimony networks based on chloroplast DNA (cpDNA) variation, and assessed population genetic parameters using nuclear microsatellites and cpDNA haplotypes. Our data provided evidence for vicariant events as well as for hybridization and cpDNA introgression among closely related Macaranga species. The extent of population differentiation within myrmecophytic versus non-myrmecophytic species proved to be in a similar range, indicating that our working hypothesis of enhanced allopatric speciation in myrmecophytes cannot be sustained by the present evidence. Nevertheless, the mutualistic interactions of Macaranga and associated ants may be a key innovation that opened an adaptive zone putatively exploited by the divergence of Macaranga.

No evidence for effects of infection with the amphibian chytrid fungus on populations of yellow-bellied toads

The parasitic chytrid fungus Batrachochytrium dendrobatidis (Bd) can cause the lethal disease chytridiomycosis in amphibians and therefore may play a role in population declines. The yellow-bellied toad Bombina variegata suffered strong declines throughout western and northwestern parts of its range and is therefore listed as highly endangered for Germany and the federal state of Hesse. Whether chytridiomycosis may play a role in the observed local declines of this strictly protected anuran species has never been tested. We investigated 19 Hessian yellow-bellied toad populations for Bd infection rates, conducted capture-mark-recapture studies in 4 of them over 2 to 3 yr, examined survival histories of recaptured infected individuals, and tested whether multi-locus heterozygosity of individuals as well as expected heterozygosity and different environmental variables of populations affect probabilities of Bd infection. Our results show high prevalence of Bd infection in Hessian yellow-bellied toad populations, but although significant decreases in 2 populations could be observed, no causative link to Bd as the reason for this can be established. Mass mortalities or obvious signs of disease in individuals were not observed. Conversely, we show that growth of Bd-infected populations is possible under favorable habitat conditions and that most infected individuals could be recaptured with improved body indices. Neither genetic diversity nor environmental variables appeared to affect Bd infection probabilities. Hence, genetically diverse amphibian specimens and populations may not automatically be less susceptible for Bd infection.