Biochemical Genetic Relationships Among Tunisian Hares (Lepus sp.), South African Cape Hares (L. capensis), and European Brown Hares (L. europaeus)

Tunisian hares (n = 45), currently assigned to Lepus capensis, were assayed for allelic variation at 40 allozyme loci, and allele frequencies at 32 loci were directly compared with earlier data of South African cape hares (L. capensis, n = 9) and European brown hares (L. europaeus, n = 244) to reveal genetic relationships among them. European mountain hares (L. timidus, n = 200) were used for outgroup comparison. In the Tunisian hares 27.5% of the loci were polymorphic with 2–4 alleles. Among all alleles at polymorphic loci, 15.1% occurred exclusively in Tunisian hares, 5.7% exclusively in cape hares, and 7.5% exclusively in brown hares at low frequencies. Not a single locus showed alternately fixed alleles between the samples of the L. capensis/L. europaeus complex. Levels of absolute and relative genetic differentiation among the samples of the L. capensis/ L. europaeus complex were low, relative to pairwise comparisons involving mountain hares. Diverse cluster analyses and multidimensional scaling of various pairwise genetic distance matrices concordantly grouped Tunisian hares with brown hares, and South African cape hares clustered only slightly farther apart, whereas mountain hares were distinctly separate. These results suggest regionally distinct phylogenetic units within an overall cohesive gene pool in the L. capensis/ L. europaeus complex, supporting Petter's view that all North African hares belong to L. capensis except for one local population of savanna hares, and that cape hares and brown hares are conspecific.     

Phylogenetic analysis of mtCR-1 sequences of Tunisian and Egyptian hares (Lepus sp. or spp., Lagomorpha) with different coat colours

North African hares are currently considered belonging to cape hares (Lepus capensis), except for an isolated occurrence of L. victoriae in NW Algeria. However, the few existing molecular data are not unequivocal. Here, we study sequence variation (415 bp) in the hypervariable domain-1 of the mitochondrial (mt) control region, of hares with different coat colour from north-central Tunisia and NW Egypt, to test Petter's [(1959): Eléments d’une révision des Lièvres africains du sous-genre Lepus. Mammalia 23, 41–67] hypothesis that North African hares belong to L. capensis. Seven Tunisian and one Egyptian haplotypes were revealed from 28 hares and compared phylogenetically to 245 haplotypes of various Lepus species downloaded from GenBank. Neighbour joining (NJ) and principal coordinate (PCO) analyses based on a Tamura-Nei 93 distance matrix, as well as maximum parsimony (MP) analysis concordantly grouped all currently obtained haplotypes together into one monophyletic clade, and revealed relatively close relationships to the clades of African scrub hares (L. saxatilis) and brown hares (L. europaeus). The three distinguished coat colour types of Tunisian hares were paralleled only to a small extent by sequence differentiation. Haplotypes of L. capensis from the nominal Cape province of South Africa, North Africa, and China clustered into different major clades, respectively, with Chinese L. capensis haplotypes forming only a subclade within a major clade that encompassed predominantly “mountain/arctic hare-type sequences” in addition to sequences of several other palaearctic and nearctic species. One further Chinese L. capensis haplotype clustered into the L. comus clade. These results indicated occurrence of introgression and/or shared ancestral polymorphism. Such an evolutionary scenario implies using nucelar markers in addition to mtDNA for phylogenetic inferences in the genus Lepus; nevertheless, mtDNA is still useful for inferring phylogenetic history and biogeography of hares.     

Mitochondrial HVI sequence variation in Anatolian hares (Lepus europaeus Pallas, 1778)

Variability and phylogenetic relationships of sequences of the hypervariable domain I (HVI) of the mitochondrial DNA was studied in 46 brown hares (Lepus europaeus) from Anatolia, to test the hypotheses that (i) hares from several islands off the Anatolian coast and from Cyprus are phylogenetically close to mainland Anatolian hares, (ii) Anatolian hare sequence variability is higher than that of typical European brown hares, and to iii) infer possible Anatolian source populations of hares from some islands in the eastern Mediterranean. Neighbor joining and Maximum Parsimony analyses revealed reciprocal monophyly for sequences from Anatolia, the considered eastern Mediterranean islands off the Anatolian coast, Cyprus, and those sequences published earlier form NE Greece that were supposed to originate from earlier immigration via the late-Pleistocene/early-Holocene land bridge that connected SE Europe and W Anatolia (Kasapidis et al., 2005. Mol. Phylogenet. Evol. 34, 55–66). A high sequence idiosyncrasy was found among the Anatolian samples. Almost all approaches to compare variability between Anatolian and the downloaded European sequence data indicated higher sequence diversity in Anatolia, in accordance with earlier findings for allozyme loci. Network and principal coordinate analyses of the Anatolian sequences and those from the islands off the Anatolian coast as well as the Anatolian-type NE Greek sequences suggested high mitochondrial gene exchange among local populations in Anatolia with little effect of possible geographic barriers, and did not provide clues for tracing possible origins of island populations.     

Biochemical genetic variability in brown hares (Lepus europaeus) from Greece

Allozyme variability of 91 brown hares (Lepus europaeus) from seven regions in Greece was compared to existing data of Bulgarian populations to test the hypothesis of the occurrence of specific alleles in Greece, likely stemming from an isolated Late Pleistocene refugial population in the southern Balkans. This hypothesis is particularly suggested by some subfossil Late Pleistocene hare remains in Greece and the reported high mtDNA diversity in Greek hares. Allozymic diversity could be higher in Greek hares than in hares from neighboring regions as a result of the accumulation of variants in a long-lasting Pleistocene refugium. Conversely, Greek hares could exhibit reduced genetic diversity because of long-lasting low effective population sizes during the Late Glacial Maximum and a lower chance of postglacial gene flow from other populations into this rather marginal part in the southern Balkans. Horizontal starch gel electrophoresis of proteins from 35~loci revealed three alleles (Es-1 ^–162, Pep-2 ¹¹⁴, Mpi ⁸⁸) at low frequencies, which were not found in Bulgarian or any other brown hare population. In contrast, some alleles from the populations from Bulgaria and other regions of Europe were absent in the Greek samples. Population genetic statistics indicated only a slight tendency of increased gene pool diversity in Greek hares, little substructuring in Greek and Bulgarian populations, respectively, as well as an only slightly lower level of gene flow between the two neighboring regions, as compared to the gene flow within each region. The results conform to the hypothesis of a Late Pleistocene refugial population in the southern Balkans, with some few specific nuclear gene pool characteristics, but little effect on the overall genetic differentiation between Greek and Bulgarian hares.     

Poxvirus fibromas on African hares.

Small dermal tumors were found on three African hares (Lepus capensis) in the Laikipia District, Kenya. Gross and histopathologic studies revealed similarities to the Shope's fibroma of wild rabbits in North America and fibromas of European hares. Histological examination of the African hare fibromas revealed intracytoplasmic inclusion bodies characteristic of poxviruses and poxvirus virions were demonstrated by electron microscopy of ultrathin sections. Attempts to propagate the virus in rabbit skin, embryonated chicken eggs and cell cultures were unsuccessful.     

Molecular evidence of conspecificity of South African hares conventionally considered Lepus capensis L., 1758

Conventionally, Lepus capensis is considered to range across large parts of Africa, the Middle East, Central and Far East Asia. However, a recent morphological study restricts cape hares tentatively to a small range in the Western Cape Region of South Africa and groups all other L. capensis-type hares from South Africa into a new species: L. centralis. Here, we studied molecular relationships among L. capensis-type hares from South Africa. Phenotypically and morphologically the individuals matched either the newly described L. capensis or L. centralis. We examined 66 hares for allelic variation at 13 microsatellite loci and for sequence variation of the hypervariable domain 1 of the mitochondrial control region. All tree-generating analyses of the currently obtained sequences and all South African cape hare sequences downloaded from GenBank revealed monophyly when compared to sequences of various other Lepus species. A network analysis indicated close evolutionary relationships between hares of the “L. capensis-phenotype” and the “L. centralis-phenotype” (according to Palacios et al. 2008) from the southwest of the Western Cape, relative to their pronounced evolutionary divergence from all other more central, northern, and north-eastern L. capensis-type hares. F-statistics, a Bayesian admixture STRUCTURE model, as well as a principal coordinate analysis of microsatellite data indicated close genetic relationships among all South African L. capensis-type hares studied presently. A coalescence model-based migration analysis for microsatellite alleles indicated gene flow between most of the considered subspecies of cape hare, including L. capensis capensis and L. capensis centralis, theoretically sufficient to balance stochastic drift effects. Concordantly, AMOVA models revealed only little effects of partitioning microsatellite variation into the two suggested morpho-species “L. capensis” and “L. centralis”. Under an “Interbreeding Species Concept” (e.g. a strict or relaxed Biological Species Concept), the current molecular data demonstrate conspecificity of the two proposed morpho-species “L. capensis” and “L. centralis”. Based on the present molecular data the differentiation of subspecies of cape hares from southern Africa is discussed.     

Differentiation under isolation and genetic structure of Sardinian hares as revealed by craniometric analysis,mitochondrial DNA and microsatellites

Hares (Lepus capensis Linnaeus 1758) were probably introduced into Sardinia in historical times. Previous studies indicated North Africa as the most likely source area but did not exclude the occurrence of hybridization events with continental brown hares (L. europaeus Pallas 1778) perhaps introduced for hunting purposes. We implemented both morphometric and genetic approaches to verify the genetic isolation of the Sardinian population. Specifically, we conducted a multivariate analysis of craniometric data and analysed 461 bp of the mitochondrial control region and 12 autosomal microsatellites in Sardinian hares, using North African cape hares and European brown hares as reference populations. Sardinian hares displayed a peculiar skull shape. In agreement, both nuclear and mitochondrial markers remarked the distinctiveness of this population. Observed and expected heterozygosity were 0.52 and 0.61, while haplotype and nucleotide diversity were 0.822 and 0.0129. Self‐assignment based on Bayesian cluster analysis was high (average membership 0.98), and no evident signs of introgression from continental brown hares were found. Our results support the hypothesis that the Sardinian hares have been introduced from North Africa, remained genetically isolated since the founding event and evolved independently from the source population. This long‐lasting isolation and the consequent genetic drift resulted in a differentiation, perhaps accompanied by an adaptation to local environmental conditions.     

Factors related to the occurrence of hybrids between brown hares Lepus europaeus and mountain hares L. timidus in Sweden

Hybridization occurs among many species, and may have implications for conservation as well as for evolution. Interspecific gene flow between brown hares Lepus europaeus and mountain hares L. timidus has been documented in Sweden and in continental Europe, and has probably to some extent occurred throughout history in sympatric areas. What local factors or ecological relationships that correlate with or trigger hybridization between these species has however been unclear. We studied spatial distribution of hybrids between brown hares and mountain hares in Sweden in relation to characteristics of the sampled localities (hunting grounds). In a sample of 70 brown hares collected from 39 populations in south‐central Sweden during 2003–2005, 11 (16%) showed introgressed mtDNA from mountain hares. Among the brown hares from their northern range, i.e. in general the most recent establishments, the corresponding figure was 75% (9/12). The frequency of samples with hybrid ancestry increased significantly with latitude, altitude and hilliness, and were higher (p<0.1) in recently established populations and/or where the proportion of arable land was low. Several site‐specific parameters were correlated, e.g. latitude as expected to hilliness, and no parameter explained the occurrence of hybrids exclusively. Instead, the appearance of mountain hare mtDNA among brown hares was associated with a conglomerate of parameters reflecting landscapes atypical for the brown hare, e.g. forest dominated and steep areas where the species quite recently was established. We suggest that these abiotic factors mirror the main aspect influencing hybridization frequency, namely the density or relative frequency of the two species. In atypical brown hare landscapes with recent establishment, mountain hares are probably relatively more common. When one species dominate in numbers, or when both species display low densities, increased frequency of hybridization is expected due to low availability of conspecific partners, a phenomenon referred to as Hubbs’ principle.     

Infections with Francisella tularensis biovar palaearctica in hares (Lepus timidus, Lepus europaeus) from Sweden

The occurrence of tularemia was studied in 1,500 hares submitted to the National Veterinary Institute, Uppsala, Sweden for postmortem examination during 1973 through 1985. A total of 109 tularemia cases was recorded based on the fluorescent antibody (FA) test for Francisella tularensis and on the gross and microscopic pathology. Tularemia was diagnosed only in the varying hare (Lepus timidus) and not in the European brown hare (Lepus europaeus). The geographical distribution of the 109 cases indicates that tularemia has not spread in Sweden during the last 45 yr, with the exception of an endemic occurrence of the disease on the island of Stora Karls? in the Baltic sea. The disease was most frequent in the autumn and only a few cases were recorded during winter. Cases were not seen in the spring. The annual prevalence varied, with several cases in 1974 and 1981, but there were no cases in 1976 and 1980. The postmortem findings in hares dying of tularemia in the autumn were characterized by focal coagulative necrosis in liver, spleen and bone marrow, with high numbers of bacteria FA-positive for F. tularensis. In hares dying during winter months, the most characteristic findings were hemorrhagic enteritis and typhlitis, although necrotic lesions could occur in liver, spleen and bone marrow. Diseased hares on the island of Stora Karls? were demonstrated to be infected with ticks, while hares on the mainland of Sweden generally were fed upon by mosquitoes. Twenty-six of the 109 hares with tularemia were examined bacteriologically and F. tularensis biovar palaearctica was isolated from eight. The lung extract antibody test for F. tularensis was performed in 18 of the 109 hares. All were negative. In addition to the field study, an experimental study with F. tularensis biovar palaearctica was performed. Four varying hares and three European brown hares were inoculated. None of the hares died from tularemia, and generalized infection was not demonstrated.     

The distribution of mountain hares Lepus timidus in Europe: a challenge from brown hares L. europaeus?

1. Throughout the most recent glacial period (Weichsel), the mountain hare Lepus timidus had a continuous distribution in the tundra habitat south of the ice‐rim. When the ice retreated, mountain hares colonized deglaciated land, and spread over northern Europe. 2. Since the Weichsel, the mountain hare's distribution in Europe has been gradually reduced and at present comprises Ireland and the Scottish Highlands, high altitudes in the Alps, isolated forests in eastern Poland, most of Fennoscandia and from the Baltic countries eastwards through Russia. Declines during the last century have been observed in Sweden and Russia. 3. This review defines and evaluates causes for this gradual reduction and fragmentation of the mountain hare's distribution, with special focus on interactions with brown hares Lepus europaeus. The relative importance of diseases, predation, cultivation and interactions with other herbivores than brown hares are discussed. 4. A plausible cause of the possible permanent disappearance of mountain hares in Europe appears to be exclusion by interspecific competition and hybridization with, and/or epidemic diseases mediated by, the congeneric brown hare.     

Speciation and paraphyly in western mediterranean hares (Lepus castroviejoi,L. europaeus,L. granatensis,andL. capensis) revealed by mitochondrial DNA phylogeny

Mitochondrial DNA (mtDNA) variation among specimens of the northwestern African hare (Lepus capensis schlumbergeri) and three European hares sampled in Spain (L. castroviejoi andL. granatensis, which are endemic to the Iberian Peninsula, andL. europaeus) was analyzed using seven restriction endonucleases. Fourteen haplotypes were found among the 34 animals examined. Restriction site maps were constructed and the phylogeny of the haplotypes was inferred. mtDNA ofL. capensis was the most divergent, which is consistent with its allopatric African distribution and with an African origin of European hares. We estimated that mtDNA in hares diverges at a rate of 1.5–1.8% per MY assuming that the European and African populations separated 5–6 MYBP. Maximum intraspecies nucleotide divergences were 1.3% inL. capensis, 2.7% inL. castroviejoi, and 2.3% inL. granatensis but 13.0% inL. europaeus. The latter species contained two main mtDNA lineages, one on the branch leading toL. castroviejoi and the other on that leading toL. granatensis. The separation of these two lineages from theL. castroviejoi orL. granatensis lineages appears to be much older than the first paleontological record ofL. europaeus in the Iberian peninsula. This suggests that the apparent polyphyly ofL. europaeus is due not to secondary introgression, but to the retention of ancestral polymorphism inL. europaeus. The results suggest thatL. europaeus either has evolved as a very large population for a long time or has been fractionated. Such a pattern of persistence of very divergent lineages has also been reported in other species of highly mobile terrestrial mammals. As far as mtDNA is concerned,L. europaeus appears to be the common phylogenetic trunk which has diversified during dispersion over the European continent and from whichL. castroviejoi andL. granatensis speciated separately in southwest Europe.     

Cytochrome b phylogeny of North American hares and jackrabbits (Lepus, lagomorpha) and the effects of saturation in outgroup taxa

Jackrabbits and hares, members of the genus Lepus, comprise over half of the species within the family Leporidae (Lagomorpha). Despite their ecological importance, potential economic impact, and worldwide distribution, the evolution of hares and jackrabbits has been poorly studied. We provide an initial phylogenetic framework for jackrabbits and hares so that explicit hypotheses about their evolution can be developed and tested. To this end, we have collected DNA sequence data from a 702-bp region of the mitochondrial cytochrome b gene and reconstructed the evolutionary history (via parsimony, neighbor joining, and maximum likelihood) of 11 species of Lepus, focusing on North American taxa. Due to problems of saturation, induced by multiple substitutions, at synonymous coding positions between the ingroup taxa and the outgroups (Oryctolagus and Sylvilagus), both rooted and unrooted trees were examined. Variation in tree topologies generated by different reconstruction methods was observed in analyses including the outgroups, but not in the analyses of unrooted ingroup networks. Apparently, substitutional saturation hindered the analyses when outgroups were considered. The trees based on the cytochrome b data indicate that the taxonomic status of some species needs to be reassessed and that species of Lepus within North America do not form a monophyletic entity.     

On shortcomings of using mtDNA sequence divergence for the systematics of hares (genus Lepus): An example from cape hares

Despite the well-known fact that evolutionary patterns of single genes or sequences are not necessarily paralleled by organismic evolution, using mitochondrial sequence divergence for inferring phylogenetic relationships among hare species is not uncommon. Earlier studies reported interspecific introgression in the genus Lepus that may slur systematic conclusions drawn exclusively from mtDNA data. We examined pairwise divergence in partial mtHV-1 sequences and nuclear gene pools based on microsatellite and allozyme loci separately in a South and North African population of cape hares, Lepus capensis, and did not find significant correspondence on the individual level within either population. Also, the former population had a significantly higher level of mtHV-1 sequence divergence compared to the latter, but levels of individual nuclear gene pool divergence did not differ significantly between the two populations. Hence, the absence of correspondence between mtHV-1 sequence divergence and nuclear gene pool divergence among individuals within a population might be independent of the population-specific level of differentiation among individuals. Our results complement earlier findings in hares, and strongly recommend to include nuclear gene pool evidence for systematic inferences within this genus, even under the absence of mitochondrial introgression.     

Evidence of autumn reproduction in female European hares (Lepus europaeus) from southern Europe

Information on reproductive biology of the European hare (Lepus europaeus) in different environmental and landscape conditions comprises part of fundamental knowledge regarding species’ adaptive responses as well as many aspects of its biology. Most of the studies conducted on European hare reproduction are confined to midlatitude and northern populations, whereas no data exist on the indigenous southern populations. Here, we present information on reproductive characteristics of European hares inhabiting Mediterranean ecosystems on the island of Crete, Greece for two successive hunting seasons. Although the annual reproductive cycle of the species is well known, with an autumn sexual inactivity, the duration of this period is subjected to fluctuations in different years and for different areas. According to our data, hare populations of Crete present an autumn–early winter reproductive activity with high proportions of pregnant females observed in all the months of the study. Furthermore, the estimated mean litter size (1.54 SE ± 0.07) while signed to the lowest values ever observed for European hares is similar to values obtained in continuous breeding species of the same genus, Lepus granatensis, Lepus corsicanus, Lepus (capensis) mediterraneus, and Lepus capensis also inhabiting warm climates. In conclusion, our results suggest that Cretan European hare populations exhibit a reproductively active period during autumn–early winter where proportions of pregnant females and litter size give a strong indication of a continuous reproduction throughout the year.     

Genetic diversity within Anatolian brown hares (Lepus europaeus Pallas, 1778) and differentiation among Anatolian and European populations

Genetic variability of Anatolian hares and relationships between Anatolian and European populations were assessed by a multilocus allozyme approach to infer evolutionary relationships between hares from Asia Minor and Europe. Of the 48 loci assayed, 19 (39.6%) were polymorphic with two to four alleles in the Anatolian hares. Among all Anatolian alleles, 14 were so far not found in the compared 717 brown hares from Europe. Overall, genetic diversity was highest in Anatolian hares, intermediate in brown hares from the southern and southeastern Balkans and lowest in central European populations. The rich genetic diversity in Anatolian hares might be a consequence of Anatolias biogeographic position with the chance of multiple gene flow from neighbouring regions, and the likelihood of long-term presence of hares during the last ice age, when large parts of more northern latitudes did not provide suitable habitats.However, among 28 loci used for the comparison between European and Anatolian populations, most common alleles of European brown hares were also common in Anatolian populations and no alternately fixed alleles were found for Anatolian and European populations. This together with only little or moderately varying allele frequencies produced low genetic divergence between Anatolian and European populations. Genetic differentiation among Anatolian populations was also low. Even between the two forms with different coat colour (brownish and yellowish) in Anatolian hares, there was little genetic differentiation. Altogether, all Anatolian hares studied presently are closely related to European brown hare populations, and only some distantly spaced population pairs revealed increased genetic divergence.

Zusammenfassung

Genetische Diversität anatolischer Feldhasen (Lepus europaeus Pallas, 1778) und Differenzierung zwischen anatolischen und europäischen PopulationenZur Beurteilung der phylogenetischen Beziehungen zwischen anatolischen Hasen und europäischen Feldhasenpopulationen wurde die allelische Variabilität anatolischer Hasen mittels horizontaler Stärkegelelektrophorese erfaßt und gemeinsam mit unmittelbar vergleichbaren Daten griechischer, bulgarischer und österreichischer Populationen aus früheren Studien populationsgenetischen Analysen unterzogen. Neunzehn der 48 untersuchten Loci der anatolischen Hasen zeigten allelische Variabilität. Unter den anatolischen Allelen kamen 14 bisher in den europäischen Polulationen nicht vor. Insgesamt zeigten anatolische Hasen die höchste und österreichische Populationen die niedrigste genetische Diversität; die jeweiligen Werte der griechischen und bulgarischen Populationen lagen dazwischen. Dies entspricht unserer Hypothese hoher genetischer Diversität in Anatolien, auf Grund der biogeografischen Position und der klimatischen bzw. Lebensraumbedingungen während des Pleistozäns, die, im Gegensatz zu Mitteleuropa, kontinuierliche Hasenpopulation in Anatolien wahrscheinlich erscheinen lassen. Kontinuierliche Populationen und Genflüsse aus verschiedenen Nachbarregionen könnten bei langfristig relative ungestörten Populationen zur Anreicherung genetischer Varianten in Anatolien geführt haben, während mitteleuropäische Feldhasenpopulationen im Zuge ihrer postglazialen Einwanderung aus Refugial-gebieten an genetischer Vielfalt eingebüßt haben. Allerdings waren die häufigen Allele der anatolischen Hasen ebenfalls häufig bei den europäischen Feldhasen vertreten; somit ergab sich insgesamt nur eine geringe genetische Differenzierung zwischen anatolischen und europäischen Feldhasen. Die zwei in Anatolien gefundenen Fellfärbungstypen (brauner vs. gelber Grundton) zeigten ebenfalls keine besondere genetische Differenzierung.     

Hares on ice: phylogeography and historical demographics of Lepus arcticus, L. othus, and L. timidus (Mammalia: Lagomorpha)

Phylogeographical investigations of arctic organisms provide spatial and temporal frameworks for interpreting the role of climate change on biotic diversity in high-latitude ecosystems. Phylogenetic analyses were conducted on 473 base pairs of the mitochondrial control region in 192 arctic hares (Lepus arcticus, Lepus othus, Lepus timidus) and two individual Lepus townsendii. The three arctic hare species are closely related. All L. othus individuals form one well-supported clade, L. arcticus individuals form two well-supported clades, and L. timidus individuals are scattered throughout the phylogeny. Arctic hare distribution was altered dramatically following post-Pleistocene recession of continental ice sheets. We tested for genetic signatures of population expansion for hare populations now found in deglaciated areas. Historical demographic estimates for 12 arctic hare populations from throughout their range indicate that L. arcticus and L. othus persisted in two separate North American arctic refugia (Beringia and High Canadian Arctic) during glacial advances of the Pleistocene, while the high genetic diversity in L. timidus likely reflects multiple Eurasian refugia.     

新疆草兔的种群遗传结构和亚种分化

新疆草兔 (Lepus capensis) 的群体遗传结构至今无系统的研究报道,亚种水平的分类也长期存在争议.该文测定了形态分类上的新疆草兔3个亚种共87个个体的线粒体DNA (mtDNA)控制区(control region,D-Loop)592 bp的序列,经分析发现148个多态性位点,共定义了44个单倍型.新疆草兔的单倍型多样度(h,0.977 ± 0.005)和核苷酸多样度(π,0.064 ± 0.031)都较高,显示了较高的遗传多样性.分子变异分析(AMOVA)结果显示,4个地理群体间的显著分化可能是由地理隔离造成的.群体遗传结构分析显示,新疆草兔包含4个进化枝,并且每个进化枝都对应特定的分布区域,显示了明显的系统地理结构.该研究的结果支持形态分类上草兔西域亚种(L.c.lehmanni)的分类地位; 但中亚亚种(L.c.centrasiaticus)被分为两个独立的进化枝,提示可能存在两个亚种; 帕米尔亚种(L.c.pamirensis)与其他亚种间的遗传距离在13%以上,提示其可能已达到种的分化水平.