Grass snakes are widely distributed across the Western Palearctic. Recent phylogeographic studies provided evidence that three distinct parapatric species exist. Two of these occur in Italy, Natrix helvetica and N. natrix, and a contact zone between both taxa has been suggested for north‐eastern Italy. Moreover, previous investigations revealed for the Italian Peninsula a complex phylogeographic structure. Using mtDNA sequences and microsatellite loci, we examined the situation for mainland Italy, Sicily, Sardinia, and Corsica. Our study confirmed the occurrence of N. natrix in north‐eastern Italy. Cline analyses revealed limited gene flow between N. helvetica and N. natrix across a narrow hybrid zone. Within N. helvetica, conflicting patterns of mitochondrial and nuclear genomic differentiation were revealed. Three nuclear genomic clusters were found; one of them corresponded to no fewer than five distinct and, in part, deeply divergent and ancient mitochondrial lineages from mainland Italy and Sicily. This cluster was paraphyletic with respect to the two remaining mitochondrial lineages, each of which matched with another nuclear genomic cluster (one from Corsica plus Sardinia and another one from western Europe north of the Alps). This unexpected pattern most likely results from mainly male‐mediated gene flow and female philopatry combined with population‐density‐dependent processes such as ‘high‐density blocking’. With respect to taxonomy, we propose to synonymize N. h. lanzai Kramer, 1970 with N. h. sicula (Cuvier, 1829), acknowledging their lacking nuclear genomic differentiation. The studied hybrid zone of N. h. helvetica and N. h. sicula in Italy is wide, with a smooth cline for nuclear markers, supporting their subspecies status. We found no evidence for the distinctiveness of the two subspecies from Corsica (N. h. corsa) and Sardinia (N. h. cetti), suggesting their synonymy, but refrain from taxonomic conclusions because of small sample sizes and the endangered status of the Sardinian taxon.
Using range‐wide sampling and 1,143 bp of mtDNA (cytochrome b gene) and 14 microsatellite loci, we examined genetic differentiation in the widely distributed Southern African angulate tortoise (Chersina angulata). We found evidence for two genealogical lineages that differ in both genetic marker systems and their preferred habitat conditions. According to a fossil‐calibrated molecular clock for all African tortoise lineages using 1,870 bp mitochondrial and 1,416 bp nuclear DNA, the two lineages of C. angulata diverged in the Pliocene (approx. 3.8 million years ago). Species distribution models reveal that the ranges of the two lineages shifted little since the Last Glacial Maximum, which is in agreement with the demographic population descriptors suggestive of stationary populations that did not experience expansion. One lineage occurs in the west, and the other in the south of the extant distribution range. In the geographic contact zone, the two lineages hybridize extensively, providing evidence for their conspecificity under the biological species concept. Each lineage could be recognized as a distinct subspecies, but the ill‐defined geographic origins of the type material of the available names prevent their identification with any taxon. With respect to the nuclear genomic markers, the western lineage shows further north‐south substructuring. A few genetically mismatched tortoises are interpreted as human translocations. Our study underlines that confiscated or captive angulate tortoises of unknown geographic provenance should not be released without prior genetic screening to avoid genetic pollution of wild populations.
In contrast to mammals, little is known about the phylogeographic structuring of widely distributed African reptile species. With the present study, we contribute data for the leopard tortoise (Stigmochelys pardalis). It ranges from the Horn of Africa southward to South Africa and westwards to southern Angola. However, its natural occurrence is disputed for some southern regions. To clarify the situation, we used mtDNA sequences and 14 microsatellite loci from 204 individuals mainly from southern Africa. Our results retrieved five mitochondrial clades; one in the south and two in the north‐west and north‐east of southern Africa, respectively, plus two distributed further north. Using microsatellites, the southern clade matched with a well‐defined southern nuclear cluster, whilst the two northern clades from southern Africa corresponded to another nuclear cluster with three subclusters. One subcluster had a western and central distribution, another occurred mostly in the north‐east, and the third in a small eastern region (Maputaland), which forms part of a biodiversity hotspot. Genetic diversity was low in the south and high in the north of our study region, particularly in the north‐east. Our results refuted that translocations influenced the genetic structure of leopard tortoises substantially. We propose that Pleistocene climatic fluctuations caused leopard tortoises to retract to distinct refugia in southern and northern regions and ascribe the high genetic diversity in the north of southern Africa to genetic structuring caused by the survival in three refuges and subsequent admixture, whereas tortoises in the south seem to have survived in only one continuous coastal refuge.
Geochelone elegans is one of the most heavily traded tortoise species of the world, and confiscated tortoises are frequently released into the wild, without knowledge about their origin. Using for the first time samples from Pakistan and Sri Lanka, we examined phylogeographic differentiation of G. elegans using 2289 bp of mitochondrial DNA. We found weak intraspecific differentiation without a clear geographic pattern. We suggest that natural phylogeographic differentiation may have been already destroyed by massive releases of confiscated non-native tortoises. The presence of two distinct clades on Sri Lanka, however, could also be the result of a natural range expansion of a mainland lineage into the distribution range of a lineage endemic to Sri Lanka during Pleistocene low sea level stands. We propose that a systematic screening of the genetic differentiation of wild G. elegans should be conducted across its entire distribution range to provide a sound basis for the relocation of confiscated tortoises.
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