From the Apennines to the Alps: colonization genetics of the naturally expanding Italian wolf (Canis lupus) population

Wolves in Italy strongly declined in the past and were confined south of the Alps since the turn of the last century, reduced in the 1970s to approximately 100 individuals surviving in two fragmented subpopulations in the central-southern Apennines. The Italian wolves are presently expanding in the Apennines, and started to recolonize the western Alps in Italy, France and Switzerland about 16 years ago. In this study, we used a population genetic approach to elucidate some aspects of the wolf recolonization process. DNA extracted from 3068 tissue and scat samples collected in the Apennines (the source populations) and in the Alps (the colony), were genotyped at 12 microsatellite loci aiming to assess (i) the strength of the bottleneck and founder effects during the onset of colonization; (ii) the rates of gene flow between source and colony; and (iii) the minimum number of colonizers that are needed to explain the genetic variability observed in the colony. We identified a total of 435 distinct wolf genotypes, which showed that wolves in the Alps: (i) have significantly lower genetic diversity (heterozygosity, allelic richness, number of private alleles) than wolves in the Apennines; (ii) are genetically distinct using pairwise F(ST) values, population assignment test and Bayesian clustering; (iii) are not in genetic equilibrium (significant bottleneck test). Spatial autocorrelations are significant among samples separated up to c. 230 km, roughly correspondent to the apparent gap in permanent wolf presence between the Alps and north Apennines. The estimated number of first-generation migrants indicates that migration has been unidirectional and male-biased, from the Apennines to the Alps, and that wolves in southern Italy did not contribute to the Alpine population. These results suggest that: (i) the Alps were colonized by a few long-range migrating wolves originating in the north Apennine subpopulation; (ii) during the colonization process there has been a moderate bottleneck; and (iii) gene flow between sources and colonies was moderate (corresponding to 1.25-2.50 wolves per generation), despite high potential for dispersal. Bottleneck simulations showed that a total of c. 8-16 effective founders are needed to explain the genetic diversity observed in the Alps. Levels of genetic diversity in the expanding Alpine wolf population, and the permanence of genetic structuring, will depend on the future rates of gene flow among distinct wolf subpopulation fragments.     

Evidence of genetic distinction and long-term population decline in wolves (Canis lupus) in the Italian Apennines

Historical information suggests the occurrence of an extensive human-caused contraction in the distribution range of wolves (Canis lupus) during the last few centuries in Europe. Wolves disappeared from the Alps in the 1920s, and thereafter continued to decline in peninsular Italy until the 1970s, when approximately 100 individuals survived, isolated in the central Apennines. In this study we performed a coalescent analysis of multilocus DNA markers to infer patterns and timing of historical population changes in wolves surviving in the Apennines. This population showed a unique mitochondrial DNA control-region haplotype, the absence of private alleles and lower heterozygosity at microsatellite loci, as compared to other wolf populations. Multivariate, clustering and Bayesian assignment procedures consistently assigned all the wolf genotypes sampled in Italy to a single group, supporting their genetic distinction. Bottleneck tests showed evidences of population decline in the Italian wolves, but not in other populations. Results of a Bayesian coalescent model indicate that wolves in Italy underwent a 100- to 1000-fold population contraction over the past 2000-10,000 years. The population decline was stronger and longer in peninsular Italy than elsewhere in Europe, suggesting that wolves have apparently been genetically isolated for thousands of generations south of the Alps. Ice caps covering the Alps at the Last Glacial Maximum (c. 18,000 years before present), and the wide expansion of the Po River, which cut the alluvial plains throughout the Holocene, might have provided effective geographical barriers to wolf dispersal. More recently, the admixture of Alpine and Apennine wolf populations could have been prevented by deforestation, which was already widespread in the fifteenth century in northern Italy. This study suggests that, despite the high potential rates of dispersal and gene flow, local wolf populations may not have mixed for long periods of time.     

How the west was won: genetic reconstruction of rapid wolf recolonization into Germany’s anthropogenic landscapes

Following massive persecution and eradication, strict legal protection facilitated a successful reestablishment of wolf packs in Germany, which has been ongoing since 2000. Here, we describe this recolonization process by mitochondrial DNA control-region sequencing, microsatellite genotyping and sex identification based on 1341 mostly non-invasively collected samples. We reconstructed the genealogy of German wolf packs between 2005 and 2015 to provide information on trends in genetic diversity, dispersal patterns and pack dynamics during the early expansion process. Our results indicate signs of a founder effect at the start of the recolonization. Genetic diversity in German wolves is moderate compared to other European wolf populations. Although dispersal among packs is male-biased in the sense that females are more philopatric, dispersal distances are similar between males and females once only dispersers are accounted for. Breeding with close relatives is regular and none of the six male wolves originating from the Italian/Alpine population reproduced. However, moderate genetic diversity and inbreeding levels of the recolonizing population are preserved by high sociality, dispersal among packs and several immigration events. Our results demonstrate an ongoing, rapid and natural wolf population expansion in an intensively used cultural landscape in Central Europe.Subject terms: Conservation biology, Population genetics     

Conservation genetics of small relic populations of silver fir (Abies alba Mill.) in the northern Apennines

Abstract

Small and isolated silver fir populations from the Emilian Apennines (northern Italy) were studied to assess their level of genetic variation and their relationship with Alpine populations. We investigated the variability of two chloroplast microsatellites to analyse the within‐population genetic variability of four peripheral and fragmented Apennine populations and to determine their phylogenetic relatedness to seven Alpine populations covering the entire distribution of silver fir in the Alps. Haplotypic richness and haplotype diversity as well as the fraction of private haplotypes were lower in Apennine populations, evidencing the genetic impoverishment of these stands. The among‐population genetic variability analysis revealed the genetic peculiarity of Apennine populations. Analysis of molecular variance showed that the highest level of the among‐population variation occurs between Alpine and Apennine regions. A neighbour‐joining dendrogram revealed a distinct Apennine cluster that included the closest Alpine population. Our genetic analysis supports a common origin for Emilian Apennine populations, suggesting that these populations are relicts of past large silver fir populations in the northern Apennines. Our results point to a relevant conservation value for these stands, to be considered in their management.     

Genetics and conservation of wolves Canis lupus in Europe

• 1 The wolf Canis lupus, the most widespread of the four species of large carnivores in Europe, after centuries of population decline and eradication, is now recovering in many countries. Wolves contribute to regulating prey–predator dynamics and interact with human activities, mainly livestock farming and ungulate hunting. Although wolves are protected in most European countries, illegal or incidental killing is widespread.
• 2 Wolf populations do not show any apparent phylogeographic structuring worldwide. Molecular and morphological studies of historical samples showed evidence of wolf ecomorph extinctions, coinciding with the great Pleistocene faunal turnover.
• 3 Extant populations show recurrent long‐range dispersal during cycles of expansion and recolonization. Demographically stable populations, in contrast, seem to be characterized by very limited gene flow.
• 4 Despite the potential for dispersal and ecological flexibility, landscape genetic approaches have demonstrated the existence of genetically distinct wolf populations, which originated through habitat and prey specializations.
• 5 Small isolated wolf populations may suffer from inbreeding depression, although selection for heterozygotes and the rescue effect can foster rapid population recovery. Population structure and dynamics is efficiently monitored by non‐invasive genetic methods, which are also useful to identify wolf × dogCanis lupus familiaris hybridization.
• 6 Despite technical advances and a better knowledge of wolf biology, wolf conservation is largely dependent on humans, and on the solution of conflicts with stakeholders.

     

Wide-range dispersal in juvenile Eagle Owls (Bubo bubo) across the European Alps calls for transnational conservation programmes

Although juvenile dispersal is an important life history component, it remains one of the less understood ecological processes regulating the dynamics of animal populations. Lack of information about patterns of dispersal hampers the estimation of the actual status and demographic trajectory of populations, and can preclude the development of sound conservation strategies. The Eagle Owl Bubo bubo is an endangered bird species in the European Alps. Many breeding sites have been abandoned in the twentieth century, although some recovery has been reported lately. Moreover, the occupancy of traditional breeding sites across years in well-monitored Alpine populations varies a lot, this despite a relatively high breeding success at the population level. This raises concern about the long-term persistence of Alpine populations. Using conventional and satellite radiotracking, we investigated the spatio-temporal dispersal of 41 juvenile Eagle Owls originating from a population in the southwestern Swiss Alps. Our main goal was to determine dispersal distances, places and times of post-dispersal settlement. Juveniles left their parents between mid-August and mid-November. They covered, on average, 12.7 km per night (linear distance between two consecutive day roosts), often crossing high mountain ranges (up to 3,000 m altitude). The mean total distance covered by an individual during dispersal was 102 km (sum of night movements), with a maximum of 230 km. Settlement places were, on average, 46 km distant from the birth place. Our study establishes long-distance dispersal in juvenile Eagle Owls, even in a complex topography, suggesting the existence of a wide-scale metapopulation system across the northwestern Alps. This metapopulation dimension should be accounted for in conservation plans.     

Distribution of genetic variability in southern populations of Scots pine (Pinus sylvestris L.) from the Alps to the Apennines

Alpine and Northern Apennine populations of Pinus sylvestris collected from eight different Italian sites were analyzed by mitochondrial nadI intron and InterSimple Sequence Repeat (ISSR) markers, in order to describe the natural level of genetic variability and to clarify their genetic relationships. The small Northern Apennine populations are the southernmost populations of this conifer in Italy. All the analyzed populations were spontaneous and reforested areas were excluded. The analysis of the polymorphisms in the nad 1 intron sequence confirmed that the Italian P. sylvestris populations have the same mitotype (mitotype a) as the Central European ones. In the genomic ISSR analysis the proportion of shared alleles between the individuals showed the highest degree of differentiation between French and Italian populations and a divergence between the Alpine and Apennine populations. Alpine populations showed a higher genetic variability (GD 0.310±0.0252) than Apennine samples (GD 0.217±0.019). In addition, the individuals from the Apennines did not show a clear population structure, suggesting a common genetic constitution of Apennine P. sylvestris. It is likely that this constitution is the result of a progressive genetic isolation between the Alpine and the Northern Apennine populations from the early Holocene. The genetic constitution of the Northern Apennine populations suggests the opportunity of a management where in situ conservation of such small populations could be coupled to their use as sources of suitable local reforesting materials.     

Dispersal Behavior and the Connectivity Between Wolf Populations in Northern Europe

ABSTRACT The isolated gray wolf (Canis lupus) population of the Scandinavian Peninsular is suffering from inbreeding depression. We studied dispersal of 35 wolves fitted with very high frequency (20) or Global Positioning System—global system for mobile (15) radiocollars in the neighboring Finnish wolf population. The growing wolf population in Finland has high numbers of dispersing individuals that could potentially disperse into the Scandinavian population. About half (53%) of the dispersing wolves moved total distances that could have reached the Scandinavian population if they had been straight-line moves, but because of the irregular pattern of movements, we detected no wolves successfully dispersing to the Scandinavian population. Dispersal to the Scandinavian population was also limited by high mortality of wolves in reindeer (Rangifer tarandus) management areas and by dispersal to Bothnian Bay at times of the year when ice was not present. We suggest that when a small wolf population is separated from source populations by distance, barriers, and human exploitation, wildlife managers could promote the population's viability by limiting harvest in the peripheral areas or by introducing wolves from the source population.     

Genetic differentiation among populations of Cicerbita alpina (L.) Wallroth (asteraceae) in the western Alps

In this study we analyzed the genetic population structure of the hygrophilous tall-herb Cicerbita alpina in the western Alps because this group of mountain plants is underrepresented in the biogeographical literature. AFLP (amplified fragment length polymorphism) fingerprints of 40 samples were analyzed from four populations situated in a transect from the southwestern Alps to the eastern part of the western Alps and one population from the Black Forest outside the Alps. Two genetic groups can be distinguished. The first group (A) comprises the populations from the northern and eastern parts of the western Alps, and the second group (B) comprises the populations from the southwestern Alps and the Black Forest. Group A originates most likely from at least one refugium in the southern piedmont regions of the Alps. This result provides molecular evidence for a humid climate at the southern margin of the Alps during the Würm glaciation. Group B originates presumably from western or northern direction and we discuss two possible scenarios for the colonization of the Alps, i.e. (1) long-distance dispersal from southwestern refugia and (2) colonization from nearby refugia in the western and/or northern Alpine forelands. The study demonstrates that the target species harbours considerable genetic diversity, even on a regional scale, and therefore is a suitable model for phylogeographic research.     

Coexistence of wolves and humans in a densely populated region (Lower Saxony,Germany)

Since the first sporadic occurrences of grey wolves (Canis lupus) west of the Polish border in 1996, wolves have shown a rapid population recovery in Germany. Wolves are known to avoid people and wolf attacks on humans are very rare worldwide. However, the subjectively perceived threat is considerable, especially as food-conditioned habituation to humans occurs sporadically. Lower Saxony (Germany) has an exceedingly higher human population density than most other regions with territorial wolves; thus, the potential for human–wolf conflicts is higher. Using hunters’ wildlife survey data from 455 municipalities and two years (2014–2015) and data from the official wolf monitoring (557 confirmed wolf presences and 500 background points) collected between 2012–2015, grey wolf habitat selection was modelled using generalized additive models with respect to human population density, road density, forest cover and roe deer density. Moreover, we tested whether habitat use changed in response to human population and road density between 2012/2013 and 2014/2015.Wolves showed a preference for areas of low road density. Human population density was less important as a covariate in the model of the survey data. Areas with higher prey abundance (5–10 roe deer/km²) and areas with >20% forest cover were preferred wolf habitats. Wolves were mostly restricted to areas with the lowest road and human population densities. However, between the two time periods, avoidance of human density decreased significantly.Recolonization of Germany is still in its early stages and it is unclear where this process will halt. To-date authorities mainly concentrate on monitoring measures. However, to avoid conflict, recolonization will require more stringent management of wolf populations and an improved information strategy for rural populations.     

Sexual differences in the behavioural response to a variation in predation risk

Predators may influence their prey populations not only through direct lethal effects, but also by causing behavioural changes. The natural expansion of the wolf (Canis lupus) into the Alps provided the rare opportunity to monitor the responses of a prey species to the return of a large predator. Density effects have rarely been considered in the study of antipredator strategies. We examined the effects of wolf recolonisation and density modifications on group size and use of safe areas by Alpine ibex (Capra ibex) in Gran Paradiso National Park (Italy), where no large terrestrial predator has been present for about a century. We documented that, in a few years, the variation in the factors affecting the landscape of fear caused significant modifications in ibex behavioural patterns that could not be accounted for by density changes only. Male groups decreased in size and moved closer to safer areas. The distance of female groups from refuge sites, instead, was not affected, and their propensity to live in groups was scarcely modified. Behavioural modifications likely caused a reduction in nutrient intake in adult male ibex, as they necessarily used lower‐quality feeding patches. Our results showed that male and female ibex, which are characterised by a strong dimorphism, adopted different strategies to solve the conflicting demands of foraging efficiently and avoiding predators.     

European common frog (Rana temporaria) recolonized Switzerland from multiple glacial refugia in northern Italy via trans‐ and circum‐Alpine routes

The high mountain ranges of Western Europe had a profound effect on the biotic recolonization of Europe from glacial refugia. The Alps present a particularly interesting case because they form an absolute barrier to dispersal for most taxa, obstructing recolonization from multiple refugia in northern Italy. Here, we investigate the effect of the European Alps on the phylogeographic history of the European common frog Rana temporaria. Based on partial cytochrome b and COXI sequences from Switzerland, we find two mitochondrial lineages roughly north and south of the Alpine ridge, with contact zones between them in eastern and western Switzerland. The northern haplogroup falls within the previously identified Western European haplogroup, while the southern haplogroup is unique to Switzerland. We find that the lineages diverged ~110 kya, at approximately the onset of the last glacial glaciation; this indicates that they are from different glacial refugia. Phylogenetic analyses suggest that the northern and southern haplogroups colonized Switzerland via trans‐ and circum‐Alpine routes from at least two separate refugia in northern Italy. Our results illustrate how a complex recolonization history of the central European Alps can arise from the semi‐permeable barrier created by high mountains.     

Southern isolation and northern long‐distance dispersal shaped the phylogeography of the widespread,but highly disjunct,European high mountain plant Artemisia eriantha (Asteraceae)

We investigated the range dynamics of Artemisia eriantha, a widespread, but rare, mountain plant with a highly disjunct distribution in the European Alpine System. We focused on testing the roles of vicariance and long‐distance dispersal in shaping the current distribution of the species. To this end, we collected AFLP and plastid DNA sequence data for 17 populations covering the entire distributional range of the species. Strong phylogeographical structure was found in both datasets. AFLP data suggested that almost all populations were genetically strongly differentiated, with 58% of the overall genetic variation partitioned among populations. Bayesian clustering identified five groups of populations: Balkans, Pyrenees, Central Apennines, one southwestern Alpine population and a Widespread cluster (eastern Pyrenees, Alps, Carpathians). Major groups were supported by neighbor‐joining and NeighbourNet analyses. Fourteen plastid haplotypes were found constituting five strongly distinct lineages: Alps plus Pyrenees, Apennines, Balkans, southern Carpathians, and a Widespread group (eastern Pyrenees, northern Carpathians, Mt. Olympus). Plastid DNA data suggested that A. eriantha colonized the European Alpine System in a westward direction. Although, in southern Europe, vicariant differentiation among the Iberian, Italian and Balkan Peninsulas predominated, thus highlighting their importance as glacial refugia for alpine species, in temperate mountain ranges, long‐distance dispersal prevailed. This study emphasizes that currently highly disjunct distributions can be shaped by both vicariance and long‐distance dispersal, although their relative importance may be geographically structured along, for instance, latitude, as in A. eriantha. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 214–226.     

Post-glacial history of the dominant alpine sedge Carex curvula in the European Alpine System inferred from nuclear and chloroplast markers

The alpine sedge Carex curvula ssp. curvula is a clonal, dominant graminoid found in the European Alps, the Carpathians, the Pyrenees and in some of the Balkan Mountains. It is a late-successional species of acidophilous alpine meadows that occurs on sites that were covered by ice during the last glacial maximum (LGM). By applying the amplified fragment length polymorphism (AFLP) fingerprinting and chloroplast DNA (cpDNA) sequencing, we attempted to identify the recolonization routes followed by the species after the last ice retreat. We relied on the genetic diversity of 37 populations covering the entire distributional range of the species. As a wind-pollinated species, C. curvula is characterized by a low level of population genetic differentiation. Nuclear and chloroplast data both support the hypothesis of a long-term separation of Eastern (Balkans and Carpathians) and Western (Alps and Pyrenees) lineages. In the Alps, a continuum of genetic depauperation from the east to the west may be related to a recolonization wave originating in the eastern-most parts of the chain, where the main glacial refugium was likely located. The Pyrenean populations are nested within the western Alps group and show a low level of genetic diversity, probably due to recent long-distance colonization. In contrast to the Alps, we found no phylogeographical structure in the Carpathians. The combination of reduced ice extension during the Würm period and the presence of large areas of siliceous substrate at suitable elevation suggest that in contrast to populations in the Alps, the species in the Carpathians underwent a local vertical migration rather than extinction and recolonization over long distance.     

Climate and habitat barriers to dispersal in the highly mobile grey wolf

We reanalysed published data to evaluate whether climate and habitat are barriers to dispersal in one of the most mobile and widely distributed mammals, the grey wolf (Canis lupus). Distance-based redundancy analysis (dbRDA) was used to examine the amount of variation in genetic distances that could be explained by an array of environmental factors, including geographical distance. Patterns in genetic variation were also examined using MDS plots among populations and relationships between genetic structure and individual environmental variables were further explored using the BIOENV procedure. We found that, contrary to a previous report, a pattern of isolation with distance is evident on a continental scale in the North American wolf population. This pattern is apparently related to climate and habitat. Specifically, vegetation types appear to play a role in the genetic dissimilarities among populations. When we controlled for the effect of spatial variation, climate was still associated with genetic distance. Further, partitioning of geographical distances into latitudinal and longitudinal axes revealed that the east-west gradient had the strongest relationship with genetic distance. We suggest two possible mechanisms by which environmental conditions may influence the dispersal decisions made by wolves.     

Genetic structure of the northwestern Russian wolf populations and gene flow between Russia and Finland

We examined the genetic diversity and structure of wolf populations in northwestern Russia. Populations in Republic of Karelia and Arkhangelsk Oblast were sampled during 1995–2000, and 43 individuals were genotyped with 10 microsatellite markers. Moreover, 118 previously genotyped wolves from the neighbouring Finnish population were used as a reference population. A relatively large amount of genetic variation was found in the Russian populations, and the Karelian wolf population tended to be slightly more polymorphic than the Arkhangelsk population. We found significant inbreeding (F = 0.094) in the Karelian, but not in the Arkhangelsk population. The effective size estimates of the Karelian wolf population based on the approximate Bayesian computation and linkage disequilibrium methods were 39.9 and 46.7 individuals, respectively. AMOVA-analysis and exact test of population differentiation suggested clear differentiation between the Karelian, Arkhangelsk and Finnish wolf populations. Indirect estimates of gene flow based on the level of population differentiation (ϕ _ST  = 0.152) and frequency of private alleles (0.029) both suggested a low level of gene flow between the populations (Nm = 1.4 and Nm = 3.7, respectively). Assignment analysis of Karelian and Finnish populations suggested an even lower number of recent migrants (less than 0.03) between populations, with a larger amount of migration from Finland to Karelia than vice versa. Our findings emphasise the role of physical obstacles and territorial behaviour in creating barriers to gene flow between populations in relatively limited geographical areas, even in large-bodied mammalian species with long-distance dispersal capabilities and an apparently continuous population structure.     

Wolves in the Great Lakes region: a phylogeographic puzzle

Empirical studies demonstrate that natural hybridization in animals is more common than thought so far ( Mallet 2005 ), particularly among species that originated recently through cycles of population contraction–expansion arising from climate changes over the last glacial period, the Pleistocene. In addition, the post‐glacial global growth of human populations has fostered anthropogenic hybridization events, mediated by habitat changes, the persecution of large predators and the introduction of alien species ( Allendorf et al. 2001 ). The Canis lineage shows cases of both natural and anthropogenic hybridization, exacerbating the controversy about the number of species that should be formally validated in the taxonomic lists, the evolutionary role of genetic introgression and the ways to manage hybrids with invading wild or domesticated populations. The study by Wheeldon et al. (2010) , published in this issue of Molecular Ecology, adds a new piece to the intricate puzzle of evolution and taxonomy of Canis in North America. They show that sympatric wolves (C. lupus) and coyotes (C. latrans) are not (extensively) hybridizing in the western North American Great Lakes region (GLR). Widespread hybridization between coyotes and a genetically distinct, but closely related, wolf‐like population (the eastern wolf) occurred in the northeastern regions of North America. In Wheeldon et al.’s (2010) opinion, these data should prove definitely that two different species of wolf (the western gray wolf C. lupus and the eastern wolf C. lycaon) and their hybrids are distributed across the GLR.     

Tyrrhenian basins of Ligury as a new peri-Mediterranean ichthyogeographic district? Population structure of <Emphasis Type="Italic">Telestes muticellus</Emphasis> (Osteichthyes,Cyprinidae), a primary freshwater fish

The phylogeographic structure of vairone (Telestes muticellus), a primary freshwater fish endangered in a large part of its distributional range, was assessed: (i) to reconstruct the complex dispersion pattern in the upper Tyrrhenian hydrographic basins of Ligury, actually not recognised as peri-Mediterranean ichthyogeographic district, and (ii) to evidence the shape of population genetic structure as useful tool for future conservation strategies. A partial fragment of mitochondrial DNA cytochrome b (497 bp) was sequenced in 109 specimens sampled from eight populations, along an east–west geographic gradient. Fourteen haplotypes were identified, confirming the evolutionary distance between the two co-generic species: T. muticellus as ‘Ligurian’ clade and T. souffia as ‘French’ clade. The Nested Clade Analysis (NCA), the population genetic variability and population structure suggested a natural colonization occurred throughout the crossing of Alpine and Apennine watershed. The hierarchical Analysis of Molecular Variance (AMOVA) confirmed a geographic distinction between the populations from western (WTL) and from eastern (ETL) Tyrrhenian basins of Ligury colonised through the river capture processes along the Maritime Alpine watershed (Padano-Venetian district) and along the Apennine watershed (Tuscano-Latium district), respectively. Our results, evidencing the lack of genetic contiguity among vairone populations of the upper Tyrrhenian hydrographic basins of Ligury, allowed to recognise the presence of two management units (MUs) for its conservation. Handling editor: Christian Sturmbauer     

中国狼不同地理种群遗传多样性及系统进化分析

为了解中国狼不同地理种群遗传多样性及系统发育情况,从中国境内狼的主要分布区青海、新疆、内蒙古和吉林4个地区采集样品,用分子生物学技术手段成功地获得44个个体线粒体DNA控制区第一高变区(HVRⅠ)序列和40个线粒体Cyt b部分序列。线粒体控制区HVRⅠ共检测到51个变异位点,位点变异率为8.76%;线粒体Cyt b部分序列发现31个变异位点,位点变异率为5.33%,未见插入及缺失现象,变异类型全部为碱基置换。共定义了16个线粒体HVRⅠ单倍型,其中吉林与内蒙种群存在共享单倍型,估计这两地间种群亲缘关系较近。4个地理种群中新疆种群拥有较高的遗传多样性(0.94)。中国狼种群总体平均核苷酸多态性为2.27%,与世界其他国家地区相比,中国狼种群拥有相对较高的遗传多样性。通过线粒体HVRⅠ单倍型构建的系统进化树可以看出,中国狼在进化上分为2大支,其中位于青藏高原的青海种群独立为一支,推测其可能长期作为独立种群进化。基于青海种群与新疆,内蒙种群的线粒体Cyt b遗传距离,推测中国狼2个世系可能在更新世冰川时期青藏高原受地质作用急速隆起后出现分歧,分歧时间大约在1.1 MY前。     

Genetic diversity and structuring of the grey wolf population from the Central Balkans based on mitochondrial DNA variation

The Dinaric-Balkan grey wolf population used to be at a border between the large remaining Eastern European populations and the largely eradicated Western European populations. During the last few decades we have witnessed the Western European wolf population recovery. Substantial genetic variation has previously been reported in the Balkan wolf population, but rigorous genetic characterization has not been done for its central parts. The aims of this research were to determine genetic diversity based on mtDNA sequence variability, to infer possible population structuring, to find genetic signals of population expansions or bottlenecks and to evaluate phylogenetic position of the grey wolf population from the Central Balkans. Six haplotypes were detected, of which three have only been found in the Balkan region. These haplotypes belong to both haplogroups previously determined in Europe. Based on our mtDNA sequence analyses, the Dinaric-Balkan wolf population is vertically differentiated into “western” (Croatia/Bosnia and Herzegovina) and “eastern” (Serbia/Macedonia) subpopulations. None of the results support assumption of population expansion. Instead, significantly positive values for Tajima's D and Fu's Fs may suggest recent population bottleneck. Obtained data may be helpful in observation to which extent gene pool from the Balkans contribute to newly founded populations in Western Europe.