Phylogeographic Differentiation Research Articles

The first European woolly rhinoceros mitogenomes, retrieved from cave hyena coprolites, suggest long-term phylogeographic differentiation.

The woolly rhinoceros (Coelodonta antiquitatis) is an iconic species of the Eurasian Pleistocene megafauna, which was abundant in Eurasia in the Pleistocene until its demise beginning approximately 10 000 years ago. Despite the early recovery of several specimens from well-known European archaeological sites, including its type specimen (Blumenbach 1799), no genomes of European populations were available so far, and all available genomic data originated exclusively from Siberian populations. Using coprolites of cave hyenas (Crocuta crocuta spelea) recovered from Middle Palaeolithic layers of two caves in Germany (Bockstein-Loch and Hohlenstein-Stadel), we isolated and enriched predator and prey DNA to assemble the first European woolly rhinoceros mitogenomes, in addition to cave hyena mitogenomes. Both coprolite samples produced copious sequences assigned to C. crocuta (27% and 59% mitogenome coverage, respectively) and woolly rhinoceros (Coelodonta antiquitatis; 27% and 81% coverage, respectively). The sequences suggested considerable DNA degradation, which may limit the conclusions to be drawn; however, the mitogenomes of European woolly rhinoceros are genetically distinct from the Siberian woolly rhinoceros, and analyses of the more complete mitogenome suggest a split of the populations potentially coinciding with the earliest fossil records of woolly rhinoceros in Europe.

Consequences of postglacial contact between phylogroups of Blarina brevicauda in North America’s Great Lakes Region

Abstract Across eastern North America, glacial cycles of the Pleistocene drove episodic latitudinal range shifts by temperate species. Isolation of populations within low-latitude refugia during glacial maxima was enhanced by physiographic barriers, leading to patterns of phylogeographic differentiation that are shared across diverse taxa. Postglacial population expansion created opportunities for differentiated lineages to come into contact, with various potential population-genetic outcomes. Northern short-tailed shrews (Blarina brevicauda) exhibit three mitochondrial phylogroups that probably originated via glacial-age range restriction and isolation. We investigate the history of postglacial expansion and interlineage contact between historically isolated regional populations of B. brevicauda. Morphological differences between skulls of shrews representing a Western lineage and those representing Central and Eastern lineages are consistent with past subspecies delineations. However, we demonstrate broad range overlap between neighboring phylogroups across the Upper Peninsula and Lower Peninsula in Michigan. Further, incongruence between phylogroup association and morphology among individuals in Upper Peninsula populations suggests that genetic admixture between shrews representing the Western and Central groups has occurred in the past and may be ongoing. We show that across most cranial measurements, shrews within the contact zone are morphologically most similar to the Central group regardless of mitochondrial identity, but one measurement in these contact zone shrews (depth of skull) is more similar to that seen in the Western group. These results suggest that hybridization between historically isolated populations has resulted in the origin of a novel skull phenotype that is proportionally deeper, narrower, and shorter than those seen in core Western and Central populations.

Cryptic diversity and phylogeographic patterns of the Bostrychia intricata species complex (Rhodomelaceae, Rhodophyta) along the coast of southeastern Australia

ABSTRACT Marine phylogeographic studies have recently shown that historical events, oceanography and ecological factors shape patterns of biodiversity and population connectivity. In southeastern Australia a historical geographic barrier, the Bassian Isthmus, results in deep genetic differentiation between some marine taxa. Using partial mitochondrial cytochrome oxidase subunit I (COI) DNA sequences, we investigated the genetic diversity and phylogeography among 26 populations of the red alga Bostrychia intricata along the southeastern Australian coast. Our genetic data revealed the occurrence of five cryptic B. intricata species (lineages N5, N7, N8, N9 and N10), two of which (N9 and N10) are newly discovered and endemic to Tasmania, while cryptic species N5 and N7 were confined to mainland Australia. Species N8 was more genetically diverse and extensively distributed than the other cryptic species, because it occurred both on mainland Australia and in Tasmania. We also detected a clear east–west phylogeographic break between cryptic species N7 and N5, and between haplotypes of species N8 on either side of Wilsons Promontory (the southernmost tip of mainland Australia). Spatial analysis of molecular variance identified three clusters of populations of species N8 along the south coast of Australia, and the split among these groups corresponded to extensive sandy areas: Coorong dunefield and Ninety Mile Beach. Additionally, our study detected the separation between mainland Australian (species N5 and N7) and Tasmanian species (N9 and N10) across the Bass Strait. We suggest that phylogeographic breaks and genetic differentiation in cryptic B. intricata species likely correlate to the historical Bassian land bridge, present-day habitats and different oceanographic circulation patterns.

Phylogeography and demographic inference of the endangered sei whale, with implications for conservation

Abstract The sei whale (Balaenoptera borealis) presents a disjointed geographic distribution. Significant genetic divergence between sei whales from the North Atlantic (NA) and North Pacific (NP) have been recorded; however, limited samples from the Southern Hemisphere (SH) have precluded a global analysis. The largest recorded mass mortality event occurred in Golfo de Penas, southern Chile, providing a primary source of SH samples. A global mitochondrial DNA phylogeographic study was undertaken including 111 SH samples (98 from Golfo de Penas), and new sequences from the NP. A marked phylogeographic structure was observed between the three ocean basins' populations, with the highest ΦST values between NA–NP and NA–SH, and a lower but still significant value between SH–NP. This structure, and the haplotype distribution pattern, suggest a more recent separation between SH–NP and an absence of recurrent gene flow among all three populations. Only one potential migratory event was detected from SH to NA. These results support an ongoing phylogeographic differentiation between all populations, highlighting the identity of each and the importance of independent management. Therefore, each population should represent a separate management unit, which is important in the development of conservation strategies. Demographic analyses detected a recent reduction of population size in the SH and NA, but not in the NP. For all populations, estimated effective population sizes were higher than estimated census sizes. This suggests that most of the genetic diversity of pre‐whaling populations is still retained; however, continued reduction in population sizes could lead to future loss of genetic diversity. This study highlights the necessity of local protection and systematic monitoring of the Golfo de Penas sei whale population. In such a remote area the use of remote sensing techniques could be a cost‐effective option for evaluating the threat, periodicity and magnitude of mortalities and their effect on the local whale population. The integration of genetic and demographic approaches improves the understanding of the dynamics of populations, which is particularly useful in endangered species.

Detection of 40bp tandem repeat motif and associated insertions and deletions (INDEL) in the mitochondrial DNA control region of Sambar deer (Rusa unicolor).

Sambar (Rusa unicolor) is the largest oriental deer and has a wide distribution across South and Southeast Asia. Despite its wide distribution range, little information on the genetics of this species is available to date. We examined the genetic variability of Sambar populations using a partial fragment of the mtDNA control region and reconstructed the phylogeographic association of Sambar populations across India. We detected 56 haplotypes with an overall haplotype diversity, Hd = 0.939 ± 0.012 and nucleotide diversity, Pi = 0.029 ± 0.002 in Sambar populations across India. The Sambar populations of Northeast India exhibits 30 localized mtDNA control region haplotypes with no haplotype sharing with other populations of India. We identified a 40bp tandem repeat motif in the mtDNA control region with variable copy numbers ranging between three to six duplications. The Sambar populations across India exhibited phylogeographic differentiation based on the observed insertion and deletion patterns in the mtDNA control region. The 40bp tandem repeat motif was associated with patterns of insertions and deletions among different populations of Sambar in India. The findings of this study provide baseline information on the mtDNA genetic variability of Sambar populations across India, which is crucial for future population genetic studies.

Population structure and gene flow of the syntopic turtles Emys and Mauremys from coastal and inland regions of Anatolia (Turkey): results from mitochondrial and microsatellite data.

Revealing the genetic basis of the existence of different species living together in different geographic regions provides clarification of this phylogeographic differentiation. In this study, we investigated the population genetics and evaluated the level of genetic variation of inland and coastal populations ofMauremys and Emys in Turkey. Tissue samples of 196 terrapins were studied which were collected fromsyntopic coastal (Gölbent-Söke/Aydın; M. rivulata and E. orbicularis) and inland populations (Bahçesaray/Aksaray; M. caspica and E. orbicularis). DNA was isolatedusing theInnuPREP DNA Mini Kit. Mitochondrial DNA sequences and allelic variation at 13 microsatellite loci for Mauremys and 12 microsatellite loci for Emys were examined. Three haplotypes were foundfor Emys orbicularis (Im, Ip and Iw) collected from the coastal region and two haplotypes for Emys orbicularis (Ig and Im) collected from inland. Two haplotypes were identified for M. caspica (Cmt8 and Cmt9) and three haplotypes were identified for M. rivulata (Rmt3, Rmt24 and Rmt26). Using microsatellites and the software STRUCTURE the most probable value for K was revealedas two 2 for both species. The FST value between M. rivulata and M. caspica was 0.39, and between the coastal and inland populations of E. orbicularis 0.09. It can be concluded that Emys populations tend to evolve by somehow preserving the allelic richness they have and Mauremys populations continue to differentiate so that new species emerge in the evolutionary process to reach the ideal allelic structure.

Genetic differentiation among populations of the blackfin goodea Goodea atripinnis (Cyprinodontiformes: Goodeidae): implications for its evolutionary history.

Central Mexico is characterized by a complex topography that is the result of historic and contemporary tectonic and climatic factors. These events have influenced the evolutionary history of numerous freshwater fishes in the region. Nonetheless, recent studies have shown that life-history traits and ecological characteristics of species may influence dispersal capabilities and the degree of genetic connectivity. Goodea (Cyprinodontiformes: Goodeidae) is one of the most widely distributed and environmentally tolerant genera of goodeids. In this study, the authors analysed variation in the mitochondrial cytochrome b gene to evaluate the phylogeographic relationships, genetic structure, genetic diversity and demographic history of Goodea from across its distribution range. They found low genetic differentiation and identified shared haplotypes among several regions. Geographic segregation was found in samples southwest and northeast of the Lower Lerma region, with some internal isolated groups showing phylogeographic differentiation and unique haplotypes. The AMOVA best explained genetic structure when grouped by haplogroups rather than when grouped by recognized biogeographic regions. Several regions showed null genetic diversity, raising the possibility of dispersal mediated by humans. Finally, Bayesian Skyline Plot analysis showed a population expansion for the Southwest haplogroup, except for the Armería population and sub-group II of the Northeast haplogroup. All this suggests a recent colonization of Goodea atripinnis throughout some of the biogeographic regions currently inhabited by this species.

Persisting in a glaciated landscape: Pleistocene microrefugia evidenced by the tree wētā Hemideina maori in central South Island, New Zealand

AbstractAimRepeated cycles of Pleistocene glaciation have influenced phylogeographic structure of taxa on New Zealand's South Island. Many taxa became restricted to refugia at either end of the island during glaciation, resulting in an area of low endemicity in central South Island. This area of low endemism is typified by the so‐called beech (or biotic) gap, where the absence of Nothofagus forest (and many other plant and invertebrate taxa) has been attributed to repeated glaciation. Some taxa, however, appear to have persisted in situ in localized refugia within the biotic gap. We test these alternative hypotheses in a large flightless alpine wētā (grasshopper).LocationSouthern Alps, South Island, New Zealand.TaxonHemideina maori Pictet & Saussure, 1891 (Orthoptera: Anostostomatidae).MethodsWe used phylogeographic analysis of mitochondrial cytochrome c oxidase I (cox1) and 25 nuclear DNA (nuDNA) markers to test for Pleistocene glacial microrefugia within the current montane South Island range of Hemideina maori.ResultsWe identified eight deeply differentiated mtDNA lineages with limited sharing of haplotypes among populations. Genetic differentiation assessed using nuDNA revealed a similar pattern, with three groups broadly corresponding to the deepest mtDNA splits. The central South Island region exhibits substantial endemic mtDNA diversity and a distinctive nuclear lineage.Main conclusionsThese results indicate that H. maori likely persisted in microrefugia within the biotic gap during glaciation. These deep lineages are estimated to have started diverging prior to the initiation of glaciation, up to 3 Ma. These results add to a growing number of Southern Hemisphere examples of deep phylogeographic differentiation in glaciated alpine regions compared to Europe and North America, probably reflecting less intense glaciation. We suggest that other Southern Alps species showing northern and southern clades alone are more montane than alpine, and were reliant on warmer habitat to the north and south during glacial eras. Thus, there are species‐specific responses to climatic processes, influenced by distinctive habitat requirements and physiological traits.

Evolution and domestication of the Bovini species.

Domestication of the Bovini species (taurine cattle, zebu, yak, river buffalo and swamp buffalo) since the early Holocene (ca. 10000 BCE) has contributed significantly to the development of human civilization. In this study, we review recent literature on the origin and phylogeny, domestication and dispersal of the three major Bos species - taurine cattle, zebu and yak - and their genetic interactions. The global dispersion of taurine and zebu cattle was accompanied by population bottlenecks, which resulted in a marked phylogeographic differentiation of the mitochondrial and Y-chromosomal DNA. The high diversity of European breeds has been shaped through isolation-by-distance, different production objectives, breed formation and the expansion of popular breeds. The overlapping and broad ranges of taurine and zebu cattle led to hybridization with each other and with other bovine species. For instance, Chinese gayal carries zebu mitochondrial DNA; several Indonesian zebu descend from zebu bull×banteng cow crossings; Tibetan cattle and yak have exchanged gene variants; and about 5% of the American bison contain taurine mtDNA. Analysis at the genomic level indicates that introgression may have played a role in environmental adaptation.

Already too late? Massive trade in Indian star tortoises (Geochelone elegans) might have wiped out its phylogeographic differentiation

Abstract 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.

Genetics of the Andean bear (Tremarctos ornatus; Ursidae, Carnivora) in Ecuador: when the Andean Cordilleras are not an Obstacle

One of the top carnivores in the Andean mountains is the Andean bear (Tremarctos ornatus, Ursidae), the only bear in South America. This is a flagship and key umbrella species in Ecuador because its conservation has a positive impact on the conservation of many other species in the Andes. But to preserve, first one must know the genetic characteristics of a species, among other things. For this, we analyzed six mitochondrial genes and seven nuclear DNA microsatellites of 108 Andean bear specimens sampled throughout Ecuador. We adopted three strategies for analyzing the data: by Province, by Region (north vs south), and by Cordillera. Four main results were obtained. First, the mitochondrial genetic diversity levels were elevated, but there were no differences in genetic diversity by Province or by Cordillera. By Regions, southern Ecuador had higher genetic diversity levels than to northern Ecuador. The genetic diversity for the microsatellites was only medium for the Andean bear at this country. Second, there was clear and significant evidence of female population expansions, for the overall sample, by Province, Region, and Cordillera. This population expansion was determined to have occurred in the time interval of 30,000–20,000 years ago (YA), during the last phase of the Pleistocene. We detected a population decrease to have occurred more recently, within the last 5000 years. It continued until about 300–200 YA when a population increase was again detected. Third, there were, practically, no phylogeographic pattern nor genetic differentiation among Andean bear populations in Ecuador by Province or by Cordillera for either mitochondrial or microsatellite markers. There was a little more genetic differentiation between northern and southern areas. Fourth, there was no trace of significant spatial genetic structure for the Andean bear in Ecuador in agreement with the genetic differentiation analyses. This shows that the Andean Cordilleras in this country did not present an obstacle to the dispersion of this species. Therefore, all of the Andean bear specimens in Ecuador should be treated as a unique Management Unit (MU) for conservation purposes, differently to that determined for other countries as Colombia.

Defining the phylogeographic relationship between cis- and trans-Andean populations of Dendrocincla fuliginosa and Xenops minutus in Colombia

Due to the position of Colombia between Central and South America, and the presence of several ecosystems in its territory, phylogeographic information from populations across this country is fundamental to understand the evolutionary history of widespread Neotropical species. The Andes have long been noted for their influence isolating lowland species into cis- and trans-Andean populations. However, detailed sampling across cis-Andean zones adjacent to the Andes (i.e., Orinoquia) has been lacking in avian phylogeographic studies. Information from DNA sequences, from 2 passerine birds: Dendrocincla fuliginosa and Xenops minutus is presented herein; to depict their phylogeographic patterns, focusing on the relationship between cis- and trans-Andean populations from Colombia. The analyses, regarding Colombian samples indicated that cis-Andean populations (i.e., Orinoquia and Amazon) are not closely related, rather the Orinoquia populations are more closely related to trans-Andean populations in both species. These relationships suggest that populations on both sides of the Andes were connected in the recent past (less than 1 myA). I propose that phylogeographic differentiation in lowland species in this region is not only explained by the presence of the Andean mountains, but also based on the ecological shifts between major ecosystems such as Amazonia and Orinoquia.

Sharp phylogeographical differentiation near the southern range edge of the silverside Odontesthes nigricans: Distinct peripheral populations and incipient speciation?

The silverside Odontesthes nigricans is the Atheriniform species that reach the southernmost latitude in Patagonia. Its wide distribution ranges from temperate waters in the Atlantic coast of South America (39°S, Buenos Aires Province, Argentina) to the Sub-Antarctic environment of Cape Horn (56°S). A previous study has shown significant intraspecific differences on growth parameters and otolith morphology between the southern peripheral population of O. nigricans in the Beagle Channel and the populations in the Atlantic coast of Tierra del Fuego. In this work, we report the first data on population genetics of O. nigricans along the southern part of its distributional range, including the peripheral populations of Beagle Channel. To examine the genetic variability, we analyzed 224 sequences of 815 bp of mitochondrial control region generated from individuals captured at two sites in the Beagle Channel and six sites at the Atlantic coast. The genetic structure assessment revealed a sharp phylogeographic division between Beagle Channel and the Atlantic coast of Patagonia with a 39.83% of variance among groups. The observed genetic diversity was lower in Beagle Channel than in the Atlantic coast sites and the demographic analyses are compatible with a scenario of past population expansion, especially in Beagle Channel. We hypothesize two possible scenarios: one is a recent colonization of the Beagle Channel from the Atlantic Coast followed by divergence promoted by adaptation to local conditions. On the other hand, population differentiation may have been attained or initiated in allopatric conditions during Quaternary glacial phases and the populations may now be experiencing a secondary contact near the eastern tip of Tierra del Fuego. These hypotheses should be addressed in the future with genomics tools. By now, these populations should be considered as different units for conservation and management purposes.

Phylogeography of the widespread creek chub Semotilus atromaculatus (Cypriniformes: Leuciscidae).

The extent and nature of genetic differentiation in Semotilus atromaculatus, one of the most abundant and widespread leuciscids in North America, were evaluated based on mitochondrial (mt) and nuclear DNA sequence variation. Phylogenetic relationships were first inferred based on a fragment of the cytochrome b (cytb) region and the nuclear intron s7 gene for S. atromaculatus and all other congeners as well as representative species from all other genera in the creek chub-plagopterin clade. The phylogeography of major haplogroups of S. atromaculatus was also assessed according to variation in a fragment of the mitochondrial cytb region from 567 individuals across its range. All analyses identified S. thoreauianus, S. lumbee and S. corporalis as reciprocally monophyletic groups. Analyses of nuclear sequence variation resolved S. atromaculatus as a single clade, where S. thoreauianus and S. lumbee were recovered as the sister group to S. atromaculatus, and S. corporalis was resolved as sister to all other species in the genus. Analyses of mtDNA sequence variation recovered S. atromaculatus as three well supported and differentiated monophyletic groups, with a widespread genetically homogeneous lineage extending across most of the current range of the species; a more geographically restricted and geographically structured lineage in the southern Appalachians, sister group to S. lumbee; and a geographically restricted lineage was identified from two Gulf Slope basins. Evidence of complex mito-nuclear discordance and phylogeographic differentiation within S. atromaculatus illustrates that further analysis of widespread species is warranted to understand North American freshwater fish diversity and distributions.

In quest of contact: phylogeography of helmeted terrapins (Pelomedusa galeata, P. subrufa sensu stricto).

Based on rangewide sampling and three mitochondrial and two nuclear markers (together up to 1,850 bp and 1,840 bp, respectively), we examine the phylogeography of two helmeted terrapin species (Pelomedusa galeata and P. subrufa sensu stricto) and infer shifts of climatically suitable spaces since the Last Glacial Maximum using a modeling approach. Whilst P. galeata displays significant phylogeographic structuring across its range and consists of two deeply divergent lineages that could represent distinct species, P. subrufa shows no obvious phylogeographic differentiation. This seems to be related to historically stable or fluctuating ranges. One of the lineages within P. galeata appears to be confined to the westernmost, winter-rainfall region of South Africa and deserves special conservational attention due to the scarcity of surface water. The other lineage is distributed further east and is differentiated in three weakly supported subclades with parapatric distribution; one occurring inland, and two along the south and east coasts, respectively. As far as is known, P. subrufa occurs in South Africa only in the northeast of the country (Limpopo, Mpumalanga) and we report the species for the first time from the Lapalala Wilderness Area in the Waterberg region (Limpopo), approximately 350 km further west than previously recorded. We confirmed the occurrence of P. galeata only 80 km south of Lapalala. Thus, a sympatric occurrence of P. galeata and P. subrufa is possible. Another putative contact zone, for the two lineages within P. galeata, must be located in the Western Cape region, and further contact zones are likely for the eastern subclades within P. galeata. The nuclear loci provided no evidence for gene flow across taxa or genetic clusters within taxa. Future investigations should use denser sampling from putative contact zones and more nuclear markers to re-examine this situation. Despite few phylogeographic studies published for southern African biota, it seems likely that differentiation follows general rules, and that climate and physiographic barriers (e.g., the Great Escarpment) have shaped phylogeographic patterns.

Evidence for panmixia despite barriers to gene flow in the hooked mussel, Ischadium recurvum (Mytilidae; Brachidontinae) along the North American coastline

The discovery of phylogeographic patterns within broadly distributed marine species can be particularly challenging because absolute physical barriers to dispersal can be inconspicuous. Genetic boundaries often lie where ocean currents meet, forming sharp physical and ecological gradients, which may act as barriers to successful migrants. In eastern North America, coastal species often show phylogeographic differentiation associated with two recognized genetic barriers: the Gulf/Atlantic and the Virginia/Carolina discontinuities. We examined 185 specimens of the intertidal hooked mussel Ischadium recurvum collected from 15 locations along the eastern coastline of North America to examine phylogeographic, migration and historical demographic patterns associated climate change associated with Pleistocene glacial patterns. Hypothesis testing using Bayes factors in Migrate-n rejected the presence of phylogeographic breaks consistent with either maritime discontinuity and favoured a panmictic population model. The migration rate from the Gulf to the Atlantic was approximately three times higher than the migration from the Atlantic to the Gulf whereas the Carolina–Virginia migration rates were nearly equal. The summary statistics (Tajima’s D, Fu’s Fs) were significant and the demographic analyses (mismatch distributions, Bayesian skyline plot) were consistent with patterns of population expansion following glacial retreat during the Pleistocene epoch.

Phylogeography of the Ibero-Maghrebian red-eyed grass snake (Natrix astreptophora)

We examined phylogeographic differentiation of the red-eyed grass snake (Natrix astreptophora) using 1984 bp of mtDNA and 13 microsatellite loci from specimens collected across its distribution range in southwestern Europe and northwestern Africa. Based on phylogenetic analyses of mtDNA, European N. astreptophora constituted the sister clade to a weakly supported North African clade comprised of two deeply divergent and well-supported clades, one corresponding to Moroccan snakes and the other to snakes from Algeria and Tunisia. This tripartite differentiation was confirmed by analyses of microsatellite loci. According to a fossil-calibrated molecular clock, European and North African N. astreptophora diverged 5.44 million years ago (mya), and the two Maghrebian clades split 4.64 mya. These dates suggest that the radiation of the three clades was initiated by the environmental changes related to the Messinian Salinity Crisis and the reflooding of the Mediterranean Basin. The differentiation of N. astreptophora, with distinct clades in the Iberian Peninsula and in the western and eastern Maghreb, corresponds to a general phylogeographic paradigm and resembles the differentiation found in another co-distributed Natrix species, the viperine snake (N. maura). Despite both species being good swimmers, the Strait of Gibraltar constitutes a significant biogeographic barrier for them. The discovery that North Africa harbours two endemic lineages of N. astreptophora necessitates more conservation efforts for these imperilled snakes.

The ‘Ward Line–Mekong–Salween Divide’ is an important floristic boundary between the eastern Himalaya and Hengduan Mountains: evidence from the phylogeographical structure of subnival herbs Marmoritis complanatum (Lamiaceae)

The Ward Line-Mekong-Salween Divide (MSD), a classic geographical barrier, is considered the main driver for population differentiation of species in the Himalaya-Hengduan Mountains region. To identify the role of geological factors in shaping the population history of subnival plant species, we investigated the phylogeography of Marmoritis complanatum based on plastid and nuclear ribosomal DNA. The phylogeographical differentiation of M. complanatum was identified between two geographical regions (central-eastern Himalaya vs. Hengduan Mountains) across the MSD, and the differentiation of two lineages began in the early Pliocene (4.49 Mya), coinciding with the formation of the MSD. These results indicate the MSD was the main driver of phylogeographical differentiation in M. complanatum. High genetic diversity and inter-population differentiation were detected in M. complanatum. Mismatch distribution analyses and species distribution modelling indicated that the broad-scale species distributions remained relatively stable with signs of a slight shrinkage of populations after the Last Glacial Maximum, suggesting Quaternary glaciations had little impact on the large-scale distributional range shift of M. complanatum. This study provides the first evidence for the effects of the MSD on the phylogeographical structure of this subnival plant species, and verifies the importance of the floristic boundary represented by the MSD.

Pattern of genetic differentiation of an incipient speciation process: The case of the high Andean killifish Orestias.

During the Pleistocene and Holocene, the southwest Andean Altiplano (17°-22°S) was affected by repeated fluctuations in water levels, high volcanic activity and major tectonic movements. In the early Holocene the humid Tauca phase shifted to the arid conditions that have lasted until the present, producing endorheic rivers, lakes, lagoons and wetlands. The endemic fish Orestias (Cyprinodontidae) represents a good model to observe the genetic differentiation that characterizes an incipient speciation process in allopatry since the morphospecies described inhabit a restricted geographic area, with present habitat fragmentation. The genetic diversity and population structure of four endemic morphospecies of Orestias (Cyprinodontidae) found in the Lauca National Park (LNP) analyzed with mitochondrial markers (Control Region) and eight microsatellites, revealed the existence of genetic groups that matches the fragmentation of these systems. High values of genetic and phylogeographic differentiation indices were observed between Chungará Lake and Piacota lagoon. The group composed of the Lauca River, Copapujo and Chuviri wetlands sampling sites showed a clear signal of expansion, with a star-like haplotype network. Levels of genetic differentiation were lower than in Chungará and Piacota, suggesting that these localities would have differentiated after the bottlenecks linked to the collapse of Parinacota volcano. The Parinacota sample showed a population signal that differed from the other localities revealing greater genetic diversity and a disperse network, presenting haplotypes shared with other LNP localities. A mixing pattern of the different genetic groups was evident using the microsatellite markers. The chronology of the vicariance events in LNP may indicate that the partition process of the Orestias populations was gradual. Considering this, and in view of the genetic results, we may conclude that the morphospecies from LNP are populations in ongoing differentiation process.

Phylogeographic differentiation versus transcriptomic adaptation to warm temperatures in Zostera marina, a globally important seagrass.

Populations distributed across a broad thermal cline are instrumental in addressing adaptation to increasing temperatures under global warming. Using a space-for-time substitution design, we tested for parallel adaptation to warm temperatures along two independent thermal clines in Zostera marina, the most widely distributed seagrass in the temperate Northern Hemisphere. A North-South pair of populations was sampled along the European and North American coasts and exposed to a simulated heatwave in a common-garden mesocosm. Transcriptomic responses under control, heat stress and recovery were recorded in 99 RNAseq libraries with ~13000 uniquely annotated, expressed genes. We corrected for phylogenetic differentiation among populations to discriminate neutral from adaptive differentiation. The two southern populations recovered faster from heat stress and showed parallel transcriptomic differentiation, as compared with northern populations. Among 2389 differentially expressed genes, 21 exceeded neutral expectations and were likely involved in parallel adaptation to warm temperatures. However, the strongest differentiation following phylogenetic correction was between the three Atlantic populations and the Mediterranean population with 128 of 4711 differentially expressed genes exceeding neutral expectations. Although adaptation to warm temperatures is expected to reduce sensitivity to heatwaves, the continued resistance of seagrass to further anthropogenic stresses may be impaired by heat-induced downregulation of genes related to photosynthesis, pathogen defence and stress tolerance.