Phylogeography of a mountain lizard species: an ancient fragmentation process mediated by riverine barriers in the Liolaemus monticola complex (Sauria: Liolaemidae)

Liolaemus monticola is a mountain lizard species, with a widespread distribution from central Chile that displays several highly polymorphic chromosomal races. Our study determined the phylogeographic structuring and relationships among three chromosomal races of L. monticola in Chile. Mitochondrial DNA (mtDNA) sequences of the cytochrome b gene were examined using the following phylogenetic methods: maximum parsimony, maximum likelihood, Bayesian inference and nested clade phylogeographic analyses (NCPAs). These methods revealed two major monophyletic clades (north and south) in the L. monticola species, with non-overlapping geographical locations separated by the Maipo and Yeso rivers (except one hybrid, from a zone of secondary contact). The NCPA showed that a past fragmentation process likely resulted in the separation of the two clades. The southern clade includes all samples of the 'Southern, 2 n  = 34' race; the northern clade is comprised of all remaining derived chromosomal races: the 'Northern, 2 n  = 38–40 and the Multiple Fission, 2 n  = 42–44' races. Our results support the hypothesis of a geographical and genetic split resulting from allopatric processes caused by riparian barriers acting over a long time period. The inferred biogeographical scenario shows that populations have moved from the south to the north using the Andean mountains as the primary corridor for dispersal.     

Reproductive isolation between chromosomal races of the house mouse Mus musculus domesticus in a parapatric contact area revealed by an analysis of multiple unlinked loci

The house mouse, Mus musculus domesticus, exhibits a high level of chromosomal polymorphism because of the occurrence and fast fixation of Robertsonian fusions between telocentric chromosomes. For this reason, it has been considered a classical speciation model to analyse the role of the chromosomal changes in reproductive isolation. In this study, we analysed a parapatric contact area between two metacentric races in central Italy, the Cittaducale race (CD: 2n = 22) and the Ancarano race (ACR: 2n = 24), to estimate gene flow at the boundary. Hybrids between these two races show high levels of structural heterozygosity and are expected to be highly infertile. A sample of 88 mice from 14 sites was used. The mice were genotyped by means of eight microsatellite loci mapped in four different autosomal arms. The results show clear genetic differentiation between the CD and ACR races, as revealed by differences in allele frequencies, factorial correspondence analysis and indexes of genetic population (e.g. F(ST) and R(ST)) along the contact zone. The genetic differentiation between the races was further highlighted by assignation and clustering analyses, in which all the individuals were correctly assigned by their genotypes to the source chromosomal race. This result is particularly interesting in view of the absence of any geographical or ecological barrier in the parapatric contact zone, which occurs within a village. In these conditions, the observed genetic separation suggests an absence of gene flow between the races. The CD-ACR contact area is a rare example of a final stage of speciation between chromosomal races of rodents because of their chromosomal incompatibility.     

Is chromosomal speciation occurring in house mice in Tunisia?

Two chromosomal races of house mice are present in Tunisia, one represented by mice carrying the 40-acrocentric standard karyotype and the other by a Robertsonian race (2re = 22) homozygous for nine centric fusions (Rb). A comparative summary on allozyme divergence, geographical distribution and level of reproductive isolation in the Tunisian and European Rb races is presented, to which new data on mitochondrial DNA and morphological divergence are added. The Tunisian 22Rb race revealed unique features not matched by the European chromosomal races, such as a decrease in allozymic variability, a higher level of genetic and morphological differentiation and a mosaic geographical distribution. The mtDNA analysis argued in favour of a local origin of the chromosomal divergence suggesting that the higher level of differentiation between the Tunisian races resulted from the older age of the 22Rb race and/or from a severe botdeneck. The decrease in fertility of chromosomal hybrids between the Tunisian races was compatible with the limited genetic introgression between diem. Furthermore, data on the restricted distribution of hybrid populations suggested that premating reproductive barriers may be evolving. The Tunisian 22Rb race is thus an appropriate model to investigate a chromosomally-mediated speciation event.     

EXTREME KARYOTYPIC VARIATION IN A MUS MUSCULUS DOMESTICUS HYBRID ZONE: THE TOBACCO MOUSE STORY REVISITED

The Robertsonian fusion is a common chromosomal mutation among mammal species and is especially prevalent in the West European house mouse, Mus musculus domesticus. More than 40 races of the house mouse exist in Europe, including the famous “tobacco mouse” (Poschiavo race) of Val Poschiavo, Switzerland. Documented here is the discovery of an extreme case of karyotypic variation in the neighboring Upper Valtellina, Italy. In a 20-km stretch of the valley, 32 karyotypes were observed, including five chromosomal races and 27 hybrid types. One previously unknown race is reported, the “Mid Valtellina” race, with a diploid number of 2n = 24 and the Robertsonian fusions Rb(1.3), Rb(4.6), Rb(5.15), Rb(7.18), Rb(8.12), Rb(9.14), Rb(11.13), and Rb(16.17). The Poschiavo race (2n = 26), Upper Valtellina race (2n = 24), Lower Valtellina race (2n = 22) and all-acrocentric race (2n = 40) were also present. The races form a patchy distribution, which we term a “mottled hybrid zone.” Geographical position, isolation, extinction, recolonization, and selection against hybrids are all believed to be instrumental in the origin and evolution of this complex system. Previous studies of house mice from Upper Valtellina indicated that two of the races in the valley (the Upper Valtellina and Poschiavo races) may have speciated in the village of Migiondo. We discuss the possibility that there may have been a reinforcement event in this village.     

Parapatric hybridization in the chromosomal speciation of the house mouse

Chromosomal races of the house mouse ( Mus domesticus ) occur extensively in Italy. The present paper reports results of studies on a hybrid zone in Central Italy on a chromosomal race with a 2 n = 22 karyotype and surrounding populations with a standard 2 n = 40 karyotype. Karyotypic and morphometric patterns of variation in this area are discussed in an attempt to contribute to an understanding of the chromosomal speciation in this species. Data are compared, where possible, with other findings on the same hybrid zone. Finally, it is suggested that changes in behavioural traits (inter-male aggression and female socio-sexual preferences) may help in promoting complete genetic isolation.     

Studies of mitochondrial DNA,allozyme and morphometric variation in a house mouse hybrid zone

An unusual chromosomal hybrid zone of the house mouse, Mus musculus domesticus, exists in Upper Valtellina, Northern Italy, consisting of four Robertsonian (Rb) races and the standard (all-acrocentric, or 2n = 40) race, all hybridizing freely within 10 km2. The hybrid zone in Valtellina provides an excellent opportunity to study the role of Rb fusions in reproductive isolation and speciation. This hybrid zone has already been well studied for the distribution of Rb fusions and the fertility of hybrids, but in order to understand the dynamics of the zone, a basic understanding of the origin and genetic similarity of the chromosomal races is necessary. This paper presents the results of three different methods of measuring genetic differentiation: multivariate analysis of morphological traits and analyses of allozyme variation and mitochondrial DNA sequences. The standard race is clearly distinguishable from the three Rb races by all three methods, but the Rb races are not distinguishable from one another. This provides strong evidence for our previous suggestions that the well-established Rb races in Valtellina are closely related, and that the standard race was introduced into the valley more recently from a distant source. The fact that the Rb races are indistinguishable is also consistent with our hypothesis that a within-village speciation event involving two of the races (Hauffe & Searle, 1992) was a recent occurrence. The low level of allozyme heterozygosity among the Rb races suggests that these populations are the products of at least one bottleneck. The present article substantially extends earlier studies and provides the first detailed morphological and molecular analysis of this complex hybrid zone.     

Mitochondrial DNA analysis confirms the existence of two glacial races of rainbow smelt Osmerus mordax and their reproductive isolation in the St Lawrence River estuary (Quebec, Canada)

A phylogeographic analysis of mitochondrial DNA variation was performed in order to test the hypothesis that north-eastern North America has been postglacially recolonized by two races of rainbow smelt Osmerus mordax . This was accomplished by documenting the geographical distribution of two major mtDNA phylogenetic clades among 1290 smelt from 49 lacustrine and anadromous populations covering most of the species' native range. The data set was built by combining previously published results with those generated in this study. The two mtDNA clades showed a geographical dichotomy, independent of life-history types, whereby the more eastern populations were either fixed or largely dominated by one clade and western populations for the other. Such geographical pattern implying a phylogenetic discontinuity provided strong evidence for the persistence of smelt in two distinct glacial refugia as well as their differential postglacial dispersal. The most likely refugium for the so-called Atlantic race was the Atlantic coastal plains, whereas that of the so-called Acadian race was the exundated Grand Banks area. Patterns of postglacial dispersal interpreted from palaeogeographic events suggested that the Atlantic race recolonized northern regions about 5000 years prior the Acadian race. Both races came into contact in the St Lawrence River estuary. While gene flow has been possible, the sympatric occurrence in the estuary of anadromous populations alternatively dominated by one mtDNA clade or the other indicated that reproductive isolation mechanisms between the two races developed within this contact zone. This represents the first evidence of secondary intergradation among distinct races of aquatic organisms in an estuarine environment.     

Electrophoretic analysis of a chromosomal hybrid zone in the grasshopper Podisma pedestris

We have made an extensive allozyme survey of 21 enzyme and protein loci in populations of the alpine grasshopper Podisma pedestris. This species occurs in two races, differing by a chromosomal fusion which separates the ancestral XO/XX race from a derived neo-XY race. These races also differ in DNA content, and hybrids between them have reduced viability. Electrophoresis reveals that the amount of genetic differentiation between these races is no greater than the variation among populations within each race. Both larger-scale surveys and a detailed survey of an area where the races hybridize, show that the chromosomal change is not correlated with gene frequency changes at any of the 21 loci studied. These findings are consistent with recently developed theory concerning the strength of the barrier to gene flow posed by a hybrid zone with characteristics such as those measured experimentally in Podisma. It is argued that hybrid zones in other species which involve allozymic differences do so because of stronger selection against hybrids rather than through mating isolation.     

A Genome Scan and Linkage Disequilibrium Analysis among Chromosomal Races of the Australian Grasshopper Vandiemenella viatica

In the past decade the interest surrounding the role of recombination in speciation has been re-kindled by a new generation of chromosomal speciation models that invoke the recombination-suppression properties of some types of chromosomal rearrangement. A common prediction of recombination-suppression models is that gene exchange between diverging populations will be more restricted in regions of the genome that experience low recombination. We carried out a genome scan of three chromosomal races of the grasshopper Vandiemenella viatica (Orthoptera: Morabinae), occurring on Kangaroo Island, South Australia, using 1517 AFLP loci, with a view to elucidating the roles that selection and chromosomal variation have played in the formation of these taxa. An analysis of molecular variance demonstrated that chromosomal race accounted for a significant proportion of the genetic variance in the total dataset, which concurred with the findings of an earlier study. Sampling across one previously-identified hybrid zone, and the identification of outlier loci between parental races allowed us to establish that, in admixed populations, outlier loci which potentially pre-date the isolation of populations of races on Kangaroo Island exhibit higher levels of linkage disequilibrium with each other than putatively neutral loci. In turn this suggests that they might reside within genomic regions of low recombination, or be closely linked with each other.     

Evidence for inversion polymorphism related to sympatric host race formation in the apple maggot fly,Rhagoletis pomonella

Evidence suggests that the apple maggot, Rhagoletis pomonella (Diptera: Tephritidae) is undergoing sympatric speciation (i.e., divergence without geographic isolation) in the process of shifting and adapting to a new host plant. Prior to the introduction of cultivated apples (Malus pumila) in North America, R. pomonella infested the fruit of native hawthorns (Crataegus spp.). However, sometime in the mid-1800s the fly formed a sympatric race on apple. The recently derived apple-infesting race shows consistent allele frequency differences from the hawthorn host race for six allozyme loci mapping to three different chromosomes. Alleles at all six of these allozymes correlate with the timing of adult eclosion, an event dependent on the duration of the overwintering pupal diapause. This timing difference differentially adapts the univoltine fly races to an approximately 3- to 4-week difference in the peak fruiting times of apple and hawthorn trees, partially reproductively isolating the host races. Here, we report finding substantial gametic disequilibrium among allozyme and complementary DNA (cDNA) markers encompassing the three chromosomal regions differentiating apple and hawthorn flies. The regions of disequilibrium extend well beyond the previously characterized six allozyme loci, covering substantial portions of chromosomes 1, 2, and 3 (haploid n = 6 in R. pomonella). Moreover, significant recombination heterogeneity and variation in gene order were observed among single-pair crosses for each of the three genomic regions, implying the existence of inversion polymorphism. We therefore have evidence that genes affecting diapause traits involved in host race formation reside within large complexes of rearranged genes. We explore whether these genomic regions (inversions) constitute coadapted gene complexes and discuss the implications of our findings for sympatric speciation in Rhagoletis.     

New Races of the Sunflower Downy Mildew Pathogen (Plasmopara halstedii) in Europe and North and South America

Collections of the causal agent of sunflower downy mildew (Plasmopara balstedii) (PH) from several European and North and South American countries were used to inoculate a series of sunflower inbred lines to differentiate races. Race 3 was identified from Argentina; race 4 from France, Hungary, and Bulgaria; race 6 from Canada and France; and race 7 from Argentina. This is the first report of race 3 in South America and of race 4 in Europe, and is the first identification of races 6 and 7. Reactions of USDA lines RHA-274, RHA-325, and DM-2 differentiate races 2, 6, and 7, with RHA-274 resistant to all three. Thirty-three commercial hybrids from 11 countries were tested; 70 % were resistant to race 2, only 27 % had resistance to race 6, and none were resistant to race 4. Surveys to determine the relative proportions of the various races within each country are needed to assess their potential impact. The USDA lines HA-335, HA-336, HA-337, HA-338, HA-339, RHA-340, HA-R4, and HA-R5 are resistant to all seven mildew races and could be utilized for the production of PH resistant hybrids. For the short term, it appears that seed treatment with metalaxyl may be the most effective control.     

Small-Scale Fragmentation Effects on Local Genetic Diversity in Two Phyllostomid Bats with Different Dispersal Abilities in Panama

Habitat fragmentation is one of the greatest threats to biodiversity. Despite their importance for conservation, the genetic consequences of small-scale habitat fragmentation for bat populations are largely unknown. In this study, we linked genetic with ecological and demographic data to assess the effects of habitat fragmentation on two species of phyllostomid bats ( Uroderma bilobatum and Carollia perspicillata ) that differ in their dispersal abilities and demographic response to fragmentation. We hypothesized that population differentiation and the effect of habitat fragmentation on levels of genetic diversity will be a function of the species' mobility. We sequenced mtDNA from 232 bats caught on 11 islands in Gatún Lake, Panamá, isolated from the mainland for ca 90 yr, and in adjacent, continuous forest on the mainland. Populations of both species showed significant genetic differentiation ( F _ST). Consistent with our prediction, population subdivision was lower in the highly mobile U. bilobatum ( F _ST= 0.01) compared to the less vagile C. perspicillata ( F _ST= 0.06), and only the latter species showed a pattern indicative of isolation by distance and, in addition, an effect of fragmentation. Genetic erosion as a result of fragmentation was also only detectable in the less mobile species, C. perspicillata , where haplotype diversity was lower in island compared to mainland populations. Our results suggest that some Neotropical bat species are prone to loss of genetic variation in response to anthropogenic small-scale habitat fragmentation. In this context, our findings point toward mobility as a good predictor of a species' vulnerability to fragmentation and altered population genetic structure.     

Contact zones between chromosomal races of Mus musculus domesticus. 3. Molecular and chromosomal evidence of restricted gene flow between the CD race (2n = 22) and the ACR race (2n = 24)

The contact zone between the Ancarano (ACR; 2n = 24) and Cittaducale (CD; 2n = 22) races of Mus musculus domesticus was studied. We used chromosomes and mitochondrial DNA (mtDNA) sequences of the control region as genetic markers to detect introgression between races. In total, 76 mice were trapped at 17 localities. Cytogenetic analysis was performed on 73 mice. A segment of the control region (468 bp) was sequenced in 41 specimens. The two races are distributed parapatrically and the contact zone was identified inside a village (Pizzoli). No mixed population was found in the study area. The contact zone does not correspond to any geographical or ecological barrier but is located in a zone of potentially high density of mice. The sequence analysis clearly demonstrates genetic differentiation between races (1.4% of sequence divergence). Hybridization is rare. Evidence of introgression was found in two individuals in the contact zone: one individual of the ACR race carries a metacentric belonging to the CD race, while another ACR individual carries a CD-like haplotype. In these ecological conditions, the observed distribution pattern and the very low level of hybridization suggest the presence of a premating mechanism of reproductive isolation.     

Distribution of genetic variability in populations of two chromosomal races of Dichroplus pratensis (Melanoplinae, Acrididae) and their hybrid zone

We examined, through allozyme electrophoresis, the genetic structure of populations of the acridid grasshopper Dichroplus pratensis from two chromosomal races (Northern and Southern) and their hybrid zone in Argentina. No fixed alleles for any particular race were found, although genetic differentiation among parental races was significant (0 = 0.044, 95% CI: 0.004-0.068). Hybrid populations are genetically more similar to the Southern race (0 = 0.008, 95% CI: -0.005-0.018) than to Northern ones (0 = 0.018, 95% CI: 0.002-0.030). Differential viability or fertility of hybrids, or asymmetry in mating preferences in favour of one particular cross would cause a higher proportion of matings between hybrid individuals and those from the Southern race. This would explain the high genetic similarity between those groups, in spite of their geographical vicinity with northern race populations.     

Interpopulation variation in fluctuating asymmetry of the palmar A-B ridge-count

Fluctuating asymmetry was measured by the Pearson correlation coefficient between a-b ridge-counts of the right and left hands. Such data were available for 38 samples, which were grouped by major geographical regions. Significant heterogeneity exists within some of the geographical groups but the between geographical group component is highly significant in both sexes. Our results show that a sizable fraction of variation in fluctuating asymmetry is related to geographical race. There is no evidence that it follows environmental or ecological lines. The variation between geographical races suggests that there is a genetic component in the magnitude of fluctuating asymmetry.     

Comparative cytogenetics of hamsters of the genus Calomyscus

Karyotypes of Calomyscus from different regions of Turkmenistan, Iran, and Azerbaijan were studied using chromosome banding (G- and C-banding) and analyses of meiosis in laboratory hybrids. Extensive variation in the diploid number and the number of autosomal arms (FNa) was revealed (2n = 30, FNa = 44; 2n = 32, FNa = 42; 2n = 44, FNa = 46; 2n = 44, FNa = 58; 2n = 37, FNa = 44; 2n = 50, FNa = 50; 2n = 52, FNa = 56). Centric and tandem fusions and heterochromatin changes were identified as the major modes of karyotype evolution in this group. Natural hybrids between individuals with different karyotypes were recorded, and regular chromosome pairing in meiosis was observed in laboratory hybrids. Fluorescence in situ hybridization with a 353-bp BspRI complex tandem repeat indicated that chromosomal repatterning occurred recently within the genus. There is no unequivocal evidence suggesting the role of chromosomal change in the speciation of the populations of Calomyscus examined.     

Hypotheses about speciation by chromosomal rearrangements in the Steropleurus martorelli complex (Tettigoniodea,Orthoptera)

Four chromosomal races ranging from 2n♂ = 29 to 2n♂ = 23 have been found in the taxonomic species S. martorelli. It is assumed that this species is not a natural entity but a species complex. Cytogenetic analysis suggests that continental race I and coastal races II, III and IV have evolved in different ways from a common ancestor called protomartorelli. The coastal races have followed a chromosome-number reduction process, arising from race II by the occurrence of two successive chromosomal fusions. The model of speciation of these coastal forms is based on chromosomal rearrangements, similar to the stasipatric model described by White, although no zones of hybridization have been found in nature.     

Mitochondrial DNA Variation and the Evolution of Robertsonian Chromosomal Races of House Mice, Mus Domesticus

The house mouse, Mus domesticus, includes many distinct Robertsonian (Rb) chromosomal races with diploid numbers from 2n = 22 to 2n = 38. Although these races are highly differentiated karyotypically, they are otherwise indistinguishable from standard karyotype (i.e., 2n = 40) mice, and consequently their evolutionary histories are not well understood. We have examined mitochondrial DNA (mtDNA) sequence variation from the control region and the ND3 gene region among 56 M. domesticus from Western Europe, including 15 Rb populations and 13 standard karyotype populations, and two individuals of the sister species, Mus musculus. mtDNA exhibited an average sequence divergence of 0.84% within M. domesticus and 3.4% between M. domesticus and M. musculus. The transition/transversion bias for the regions sequenced is 5.7:1, and the overall rate of sequence evolution is approximately 10% divergence per million years. The amount of mtDNA variation was as great among different Rb races as among different populations of standard karyotype mice, suggesting that different Rb races do not derive from a single recent maternal lineage. Phylogenetic analysis of the mtDNA sequences resulted in a parsimony tree which contained six major clades. Each of these clades contained both Rb and standard karyotype mice, consistent with the hypothesis that Rb races have arisen independently multiple times. Discordance between phylogeny and geography was attributable to ancestral polymorphism as a consequence of the recent colonization of Western Europe by mice. Two major mtDNA lineages were geographically localized and contained both Rb and standard karyotype mice. The age of these lineages suggests that mice have moved into Europe only within the last 10,000 years and that Rb populations in different geographic regions arose during this time.