Dasineura ingeris sp.n. (Diptera: Cecidomyiidae) on Salix viminalis in Sweden, including comparisons with some other Dasineura species on Salix

Infestations of an undescribed gall midge species were discovered in Southern Sweden in biomass plantations and nurseries of Salix viminalis L. Terminal leaf buds are damaged and side shoots subsequently develop. This midge, Dasineura ingeris sp.n., closely resembles three other species occurring on Salix: D.terminalis (H. Loew) on S.alba L. and S.fragilis L.; D.iteobia (Kieffer) recorded from S.caprea L. and S.cinerea L.; and D.schreiteri (Stelter) (= comb.n. for Rabdophaga schreiteri Stelter) originally found as an inquiline in the galls of another gall midge, Dasineura rosaria (H. Loew), on S.repens L. No qualitative differences in morphology were observed between these four gall midge species, but results of morphometric analyses show significant differences between all of them. In oviposition preference trials, which included host plants of all four midge species, D.ingeris laid eggs mainly on S. viminalis, D.schreiteri preferably on S.repens , and D.iteobia exclusively on S.caprea. In larval performance trials D.ingeris produced many galls on S. viminalis , one gall also on S.caprea , but no galls on either S.alba or S.repens.     

Host–pathogen relationship between Salix and Melampsora sheds light on the parentage of some biomass willows

The association between willow ( Salix ) and rust ( Melampsora ) is highly specific. Willows named Salix burjatica , S. dasyclados ( S. × dasyclados ) and S. × calodendron are important in renewable-energy plantations in the UK and western Europe. There has been much controversy over their origin, species status and nomenclature. It has been suggested that they have originated from hybridization between. S. caprea , S. viminalis and S. cinerea . In the present work, 59 willow clones were investigated through morphological examination and detached leaf inoculation using willow differentials, for their association, in southwest England, with M. capraearum and three pathotypes of Melampsora epitea (Me-A, B and C). M. capraearum was found on all clones of S. caprea and its hybrids with S. aurita ; Me-A on all S. viminalis clones; Me-B on wild S. cinerea , S. × calodendron , S. × dasyclados 'De Biardii 445' and S . 'Spaethii'; Me-C on all S. burjatica clones and most S. × dasyclados clones. Both M. caprearum and Me-A infected all S. × sericans ( S. caprea × viminalis ) clones and S. × dasyclados 'LA041/03'. We suggest that S. × dasyclados 'LA041/03' should be treated as S. × sericans ( S. caprea × S. viminalis ); S. burjatica , S. dasyclados and S. × dasyclados as synonyms; S. × dasyclados 'De Biardii 445' as S. × calodendron 'De Biardii 445'; and S . 'Spaethii' as S. × calodendron 'Spaethii'.     

Phenotypic trade-offs in the sexual reproduction of Salicaceae from flood plains

We studied the relationship of seed mass to seed longevity (controlled conditions) and to seed number in six species of Salicaceae (Populus nigra, Salix alba, S. daphnoides, S. elaeagnos, S. purpurea, and S. triandra) that frequently co-occur on European flood plains. These species regenerate sexually in the same habitat but differ in seed mass. Half-viability periods, i.e., the time after which 50% of the initially viable seeds no longer germinate, were short (between 6.5 ± 0.1 and 23.3 ± 0.3 d), and large numbers of seeds were produced (between 10?000 and 1 × 10(6) per plant). Mean seed mass ranged from 0.02 ± 0.001 mg in S. triandra to 0.80 ± 0.05 mg in P. nigra. Whereas seed mass was, against expectation, positively related to half-viability periods, seed number generally decreased with increasing seed mass. Thus, a phenotypic trade-off between seed mass and seed number appears to be accentuated by an increase in seed longevity with increasing seed mass.     

Patterns of interspecific associations of stem gallers on willows

Abstract.  The pattern of interspecific associations of three stem-galling sawfly species ( Euura atra , E. elaeagnos , E. purpureae ) and three stem-galling gallmidge species ( Rabdophaga sp. 3–5) was investigated on five willow taxa ( Salix alba , S. fragilis , S.  ×  rubens , S. elaeagnos , S. purpurea ) at five natural sites in Central Europe. The willow species harboured specific species associations of two stem gallers, each pair consisting of one Euura and one Rabdophaga species. The stem gallers were patchily distributed and their densities varied significantly among willow host plant species, host plant individuals, and host plant sexes. Four of the six species showed a significant increase in galling rate with shoot length. The other two species were the sawfly and cecidomyiid pair that induce galls on S. purpurea . The preference of stem gallers to longer shoots was generally not related to higher larval performance in terms of survival. Only one species, Rabdophaga sp. 5, was found to be more abundant on male plants. The correlation of densities of the species pairs of stem gallers was independent of willow sexes. Species pairs of stem gallers co-occurring on the same willow species tended to attack different shoots within the same host plant individual. When species pairs co-occurred on shoots they were usually found in similar densities as when occurring alone on shoots. The stem-galling sawflies usually formed galls at the basal part of a shoot, whereas the gallmidge R . sp. 5 ( R . sp. 3 and R . sp. 4 showed no clear tendency) preferred the middle or distal part of a shoot. This is interpreted with differences of their phenology and oviposition period.     

Brittleness of twig bases in the genus Salix: fracture mechanics and ecological relevance

The twig bases within the genus Salix were investigated. Brittleness of twig bases as defined in the literature neither correlates with Young's modulus nor with growth strains, which were measured for S. alba, S. fragilis and S. x rubens. For the species S. alba, S. appendiculata, S. eleagnos, S. fragilis, S. purpurea, S. triandra, S. viminalis, and S. x rubens, fracture surfaces of broken twigs were investigated and semiquantitatively described in terms of 'relative roughness' (ratio of rough area of fracture surface over whole area of fracture surface). The relative roughness clearly corresponds with the classification into brittle and nonbrittle species given in the literature. An attempt was made to quantify brittleness with mechanical tests. The absolute values of stress and strain do not correlate with the brittleness of the twig bases as defined by the relative roughness. However, the 'index stress' (ratio of stress at yield over stress at fracture) or the 'index strain' (ratio of strain at yield over strain at fracture), correlate well with the relative roughness. The graphic analysis of index stress against index strain reveals a straight line on which the eight species are ordered according to their brittleness. Depending on growth form and habitat, brittle twig bases of willows may function ecologically as mechanical safety mechanisms and, additionally, as a propagation mechanism.     

Interactions between vegetation development and island formation in the Alpine river Tagliamento

Abstract. Early-successional stages of woody vegetation on gravel bars were studied in an island-braided section of the River Tagliamento in northeastern Italy. We mapped landscape-level changes in the study area (125 ha) by GIS-based analysis of aerial photographs for two time periods (1984–1986, 1986–1991); we surveyed island vegetation, and estimated island age by tree ring analysis. The study area experienced considerable changes between 1984 and 1991 due to at least two major floods in 1987 and 1990. The development of woody vegetation on bars follows three distinct phases: (1) gravel bars plus large woody debris (LWD), (2) pioneer islands, and (3) established islands. Established islands have sections dominated by shrubs of Salix elaeagnos, S. purpurea, S. daphnoides and S. triandra, and tree-dominated sections with Populus nigra, Salix alba and Alnus incana. Large woody debris seems to play a key role for plant colonization on gravel bars. The succession from bars to established islands took about 10 - 20 yr, and the probability of an island being washed away decreased with island age. Erosion produced new LWD which again initiated successional processes in the active zone of the river. Most species were already present in the early-successional stages, although the number of species increased with island development. Established islands were characterized by a distinctive species composition, including an assemblage of species less tolerant of inundation. The results are discussed within the framework of island dynamics and its significance for restoration of early-successional habitats in more regulated rivers.     

Chloroplast DNA variation and postglacial recolonization of common ash (Fraxinus excelsior L.) in Europe

We used chloroplast polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) and chloroplast microsatellites to assess the structure of genetic variation and postglacial history across the entire natural range of the common ash (Fraxinus excelsior L.), a broad-leaved wind-pollinated and wind-dispersed European forest tree. A low level of polymorphism was observed, with only 12 haplotypes at four polymorphic microsatellites in 201 populations, and two PCR-RFLP haplotypes in a subset of 62 populations. The clear geographical pattern displayed by the five most common haplotypes was in agreement with glacial refugia for ash being located in Iberia, Italy, the eastern Alps and the Balkan Peninsula, as had been suggested from fossil pollen data. A low chloroplast DNA mutation rate, a low effective population size in glacial refugia related to ash's life history traits, as well as features of postglacial expansion were put forward to explain the low level of polymorphism. Differentiation among populations was high (GST= 0.89), reflecting poor mixing among recolonizing lineages. Therefore, the responsible factor for the highly homogeneous genetic pattern previously identified at nuclear microsatellites throughout western and central Europe (Heuertz et al. 2004) must have been efficient postglacial pollen flow. Further comparison of variation patterns at both marker systems revealed that nuclear microsatellites identified complex differentiation patterns in south-eastern Europe which remained undetected with chloroplast microsatellites. The results suggest that data from different markers should be combined in order to capture the most important genetic patterns in a species.     

A comparison of RAPD versus microsatellite DNA markers in population studies of the massasauga rattlesnake

We compared genetic differentiation among populations of the threatened massasauga rattlesnake (Sistrurus c. catenatus) using two types of nuclear molecular markers: randomly amplified polymorphic DNA (RAPD) markers and microsatellites. Analyses of molecular variance (AMOVA) and G(ST) and F(ST) analyses indicated that levels of among-population differentiation between regional populations (>100 km) were comparable for both markers. However, microsatellites were superior in population assignment tests and at discerning fine-scale genetic differentiation between subpopulations separated by tens of kilometers. These results argue that both types of markers are suitable for defining broad-scale genetic structures in snake populations and can provide important inputs into conservation initiatives of focal taxa. However, our analyses suggest that microsatellites 3re better for detecting structure at limited spatial scales.     

Camouflage and selective predation in caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata)

Final instar caterpillars of the poplar hawkmoth ( Laothoe populi ) are usually yellow-green when they have been feeding on Salix fragilis but white when on Populus alba. Similarly final instar caterpillars of the eyed hawkmoth (Smerinthus ocellata) vary in colour from yellow-green when fed on Salix cinerea or S. fragilis to grey-green when fed on Salix ripens or S. alba. Most caterpillars of these two moths are thus very cryptic to the human eye. It has already been shown that colour matching is brought about by young caterpillars changing colour depending on the colour of the substrate perceived by their eyes. Evidence is presented here for selective predation, probably by birds. Yellow-green poplar hawk caterpillars disappeared from P. alba more rapidly than did white caterpillars, but there was no significant difference in the survival of the two colour forms on S. fragilis. Yellow-green caterpillars survived better on S. fragilis than on P. alba , but there was no significant difference in the survival of white caterpillars on the two plants. Reasons why selective predation did not occur in all of the experiments are discussed.     

Autosomal, mtDNA, and Y-chromosome diversity in Amerinds: pre- and post-Columbian patterns of gene flow in South America

To evaluate sex-specific differences in gene flow between Native American populations from South America and between those populations and recent immigrants to the New World, we examined the genetic diversity at uni- and biparental genetic markers of five Native American populations from Colombia and in published surveys from native South Americans. The Colombian populations were typed for five polymorphisms in mtDNA, five restriction sites in the beta-globin gene cluster, the DQA1 gene, and nine autosomal microsatellites. Elsewhere, we published results for seven Y-chromosome microsatellites in the same populations. Autosomal polymorphisms showed a mean G(ST) of 6.8%, in agreement with extensive classical marker studies of South American populations. MtDNA and Y-chromosome markers resulted in G(ST) values of 0.18 and 0.165, respectively. When only Y chromosomes of confirmed Amerind origin were used in the calculations (as defined by the presence of allele T at locus DYS199), G(ST) increased to 0.22. G(ST) values calculated from published data for other South American natives were 0.3 and 0.29 for mtDNA and Amerind Y chromosomes, respectively. The concordance of these estimates does not support an important difference in migration rates between the sexes throughout the history of South Amerinds. Admixture analysis of the Colombian populations suggests an asymmetric pattern of mating involving mostly immigrant men and native women.     

Vole preference for Salix caprea, S. repens, and their F1, F2, and backcross hybrids

In recent years, the influence of plant hybridization on plant–animal interactions has become an important issue; however, these studies have mainly focused on insects or slugs and to a large degree ignored mammalian herbivores despite their importance in many ecosystems. This study aims to determine the preference of voles for pure Salix caprea , S. repens , and their F1, F2, and backcross hybrids to evaluate whether voles select against hybrids in natural willow populations. To address this, we conducted two field studies and a cafeteria experiment in the laboratory with bank voles ( Clethrionomys glareolus ). We found no strong indications of reduced resistance in any of the hybrid taxa. Voles damaged more S. repens than S. caprea individuals. There was a general decreasing trend of utilization from pure S. repens to pure S. caprea , hybrid taxa being intermediate between the parents. Thus, voles seemingly do not have a more detrimental effect on hybrid fitness than on the fitness of pure individuals and probably will not select against hybrids in these willows populations.     

Genetic structure and reproduction dynamics of Salix reinii during primary succession on Mount Fuji,as revealed by nuclear and chloroplast microsatellite analysis

The early stage of volcanic desert succession is underway on the southeastern slope of Mount Fuji. We used markers of nuclear microsatellites (simple sequence repeats; SSR) and chloroplast microsatellites (cpSSR) to investigate the population genetic structure and reproduction dynamics of Salix reinii, one of the dominant pioneer shrubs in this area. The number of S. reinii genets in a patch and the area of the largest genet within the patch increased with patch area, suggesting that both clonal growth and seedling recruitment are involved in the reproduction dynamics of S. reinii. Five polymorphic cpSSR markers were developed for S. reinii by sequencing the noncoding regions between universal sequences in the chloroplast genome. Nineteen different cpSSR haplotypes were identified, indicating that S. reinii pioneer genets were created by the long-distance dispersal of seeds originating from different mother genets around the study site, where all vegetation was destroyed during the last eruption. Furthermore, the clustered distributions of different haplotypes within each patch or plot suggested that newly colonized genets tended to be generated from seeds dispersed near the initially established mother genets. These results revealed that the establishment of the S. reinii population on the southeastern slope of Mount Fuji involved two sequential modes of seed dispersal: long-distance dispersal followed by short-distance dispersal.     

Phylogeography and genetic structure of disjunct Salix arbutifolia populations in Japan

Disjunct geographic distributions of boreal plant species at the southern edges of their ranges are expected to result in low genetic diversity and high genetic differentiation in the disjunct populations. This prediction was tested in a riparian willow, Salix arbutifolia, distributed in the northeastern Eurasian continent and the Sakhalin, Hokkaido, and Honshu Islands, using chloroplast DNA haplotypes and nuclear microsatellite genotypes. Hokkaido and Honshu populations shared a chloroplast haplotype identical to a closely related species, S. cardiophylla. This haplotype was divergent from haplotypes in the Eurasian continent (Primorsky) and the Sakhalin Island. In the nuclear microsatellites, most Hokkaido populations were genetically closer to Primorsky populations than to Sakhalin populations in spite of the geographical vicinity between Sakhalin and Hokkaido. The unexpected genetic divergence between Sakhalin and Hokkaido implies a complicated history of migration and colonization. The most peripheral populations in Honshu had the lowest genetic diversity and were most differentiated from the others. Thus, low genetic diversity and high genetic differentiation at the range periphery suggest substantial effects of genetic drift on genetic structure in the disjunct populations of Salix arbutifolia at the southern edge of its range.     

Chloroplast DNA phylogeography of European ashes, Fraxinus sp. (Oleaceae): roles of hybridization and life history traits

We investigated range-wide phylogeographic variation in three European ash species (Fraxinus sp., Oleaceae). Chloroplast DNA (cpDNA) microsatellites were typed in the thermophilous Fraxinus angustifolia and Fraxinus ornus and the observed haplotypes and the geographic distribution of diversity were compared to cpDNA data previously obtained in the more cold-tolerant Fraxinus excelsior. We found wide-ranging haplotype sharing between the phylogenetically close F. angustifolia and F. excelsior, suggesting hybridization (i) in common glacial refuges in the Iberian Peninsula, northern Italy, the eastern and/or Dinaric Alps and the Balkan Peninsula, and/or (ii) during postglacial recolonization. The data allowed us to propose additional glacial refuges for F. angustifolia in southern Italy and in Turkey, and populations from the latter region were particularly polymorphic. There was evidence for refuge areas in Italy, the Balkan Peninsula and Turkey for F. ornus, which did not share any single chloroplast haplotype with the other species. In both F. angustifolia and F. ornus, cpDNA diversity (h(S) = 0.027 and h(S) = 0.009, respectively) was lower and fixation levels (G(ST) = 0.964 and G(ST) = 0.983, respectively) higher than in sympatric F. excelsior (h(S) = 0.096, G(ST) = 0.870). These diversity patterns could be due to temperature tolerance or the demographic history.     

Past climate changes explain the phylogeography of Vitellaria paradoxa over Africa

The evolution of the savanna biome has been deeply marked by repeated contraction/expansion phases due to climate perturbations during the Quaternary period. In this study, we investigated the impact of the last glacial maximum (LGM) on the present genetic pattern of Vitellaria paradoxa (shea tree), a major African savanna tree. A range-wide sampling of the species enabled us to sample 374 individuals from 71 populations distributed throughout sub-Sahelian Africa. Trees were genotyped using 3 chloroplasts and 12 nuclear microsatellites, and were sequenced for 2 polymorphic chloroplast intergenic spacers. Analyses of genetic diversity and structure were based on frequency-based and Bayesian methods. Potential distributions of V. paradoxa at present, during the LGM and the last interglacial period, were examined using DIVA-GIS ecological niche modelling (ENM). Haplotypic and allelic richness varied significantly across the range according to chloroplast and nuclear microsatellites, which pointed to higher diversity in West Africa. A high but contrasted level of differentiation was revealed among populations with a clear phylogeographic signal, with both nuclear (F(ST) = 0.21; R(ST) = 0.28; R(ST) > R(ST) (permuted)) and chloroplast simple sequence repeats (SSRs) (G(ST) = 0.81; N(ST) = 0.90; N(ST) > N(ST) (permuted)). We identified a strong geographically related structure separating western and eastern populations, and a substructure in the eastern part of the area consistent with subspecies distinction. Using ENM, we deduced that perturbations during the LGM fragmented the potential eastern distribution of shea tree, but not its distribution in West Africa. Our main results suggest that climate variations are the major factor explaining the genetic pattern of V. paradoxa.     

The food plant preferences of Phratora vitellinae (Coleoptera: Chrysomelinae)

The specialisation of the beetles from the Petite Camargue on Salix nigricans was confirmed by laboratory food-plant trials. When S. nigricans is not present in the choice offered, other Salix species are accepted and the following ranking, in decreasing order of preference, can be established: S. nigricans, S. purpurea, Populus nigra, P. tremula, S. alba, s. caprea and S. cinerea. S. nigricans leaves are the richest in salicin of the 4 most acceptable plant species. The 3 least acceptable species have the undersurface of their leaves covered with trichomes.The food-plant preferences of 2 populations from central Europe and 2 from Belgium differ both in the field and in the laboratory. The level of dietary specialisation also differs from population to population. The populations originating from localities (Oignie in Belgium and the Petite Camargue in central Europe) where their favorite food-plants in the field and in the laboratory (respectively P. tremula and S. nigricans) are abundant, show clearer preferences than the populations from localities (Grammont in Belgium and Herrliberg in central Europe) where these plants are scarcer or even absent, due to human influence.The Salicaceae tested, as seen by the more specialised beetles of Oignie and the Petite Camargue, include both a strongly preferred plant (P. tremula or S. nigricans) and two or three strongly disliked species (S. alba, S. caprea and S. cinerea). for these beetles, laboratory preferences and field observations of host plant species are in agreement. It is interesting that S. nigricans, although not present in Belgium, is the second most preferred food of the Belgian beetles in laboratory trials. Similarly, P. tremula, the preferred food of the Belgian beetles, is well accepted by the central European beetles.For the less specialised beetles of Grammont and Herrliberg, the same Salicaceae include strongly disliked speicies but no single strongly preferred species. In the Herrliberg population, with rather poorly defined preferences, preference for the favorite species of the more specific population from the same geographical area (Petite Camargue) can readily be produced in the laboratory by conditioning.     

Population genetic structure of native versus naturalized sympatric shrub willows (Salix; Salicaceae)

Vegetative propagation of an introduced species can contribute significantly to its ability to spread and become naturalized, potentially in competition with native species. This study focused on the naturalization of a willow shrub, Salix purpurea, which was introduced to the United States from Europe and is commonly sympatric with the native shrub willow, S. eriocephala. Both species are capable of vegetative and sexual reproduction, but little is known about their relative frequency, nor the impact of clonal propagation on population-level genetic diversity. We analyzed genotypes at several microsatellite loci in 993 individuals belonging to 30 subpopulations of S. eriocephala and 28 subpopulations of S. purpurea in areas of sympatry across three watersheds to compare their genetic diversity and genetic structure. Our results revealed six subpopulations of S. purpurea containing plants with identical multilocus genotypes, while clonal individuals were rare among S. eriocephala populations. These species are dioecious with relatively high levels of heterozygosity, but S. eriocephala had much higher allelic diversity and genotypic diversity than did S. purpurea. These results strongly suggest that vegetative propagation has contributed to the naturalization of S. purpurea and has resulted in higher levels of genetic differentiation among S. purpurea populations than among native S. eriocephala populations.     

PERMANENT GENETIC RESOURCES: Consensus primers of cyp73 genes discriminate willow species and hybrids (Salix, Salicaceae)

Consensus primers, based on exon sequences of the cyp73 gene family coding for cinnamate 4-hydroxylase (C4H) of the lignin biosynthesis pathway, were designed for the tetraploid willow species Salix alba and Salix fragilis. Diagnostic alleles at species level were observed among introns of three cyp73 genes and allowed unambiguous detection of the first generation and introgressed hybrids in populations. Progeny analysis of a female S. alba with a male introgressed hybrid confirmed the codominant inheritance of each intron. Sequences of the diagnostic alleles of both species were similar to those found in the hybrids.     

Genome scanning for interspecific differentiation between two closely related oak species [Quercus robur L. and Q. petraea (Matt.) Liebl.

Interspecific differentiation values (G(ST)) between two closely related oak species (Quercus petraea and Q. robur) were compiled across different studies with the aim to explore the distribution of differentiation at the genome level. The study was based on a total set of 389 markers (isozymes, AFLPs, SCARs, microsatellites, and SNPs) for which allelic frequencies were estimated in pairs of populations sampled throughout the sympatric distribution of the two species. The overall distribution of G(ST) values followed an L-shaped curve with most markers exhibiting low species differentiation (G(ST) < 0.01) and only a few loci reaching >10% levels. Twelve percent of the loci exhibited significant G(ST) deviations to neutral expectations, suggesting that selection contributed to species divergence. Coding regions expressed higher differentiation than noncoding regions. Among the 389 markers, 158 could be mapped on the 12 linkage groups of the existing Q. robur genetic map. Outlier loci with large G(ST) values were distributed over 9 linkage groups. One cluster of three outlier loci was found within 0.51 cM; but significant autocorrelation of G(ST) was observed at distances <2 cM. The size and distribution of genomic regions involved in species divergence are discussed in reference to hitchhiking effects and disruptive selection.     

Phylogeographic history of white spruce during the last glacial maximum: uncovering cryptic refugia

Although a recent study of white spruce using chloroplast DNA uncovered the presence of a glacial refuge in Alaska, chloroplast failed to provide information on the number or specific localities of refugia. Recent studies have demonstrated the utility of nuclear microsatellites to refine insights into postglacial histories. The greater relative rate of mutation may allow finer scale resolution of historic dynamics, including the number, location, and sizes of refugia. Genetic data were acquired from screening 6 microsatellite loci on approximately 14 trees from each of 22 populations located across the central and western boreal forests of Canada and Alaska. Our studies combining microsatellites with Bayesian analyses of population structure in white spruce support the phylogeographic patterns uncovered using chloroplast, separating Alaskan from non-Alaskan regions. Results also support the idea that north-central Alaska served as a glacial refugium during the last glacial maximum. Additionally, the relationship between the degree of genetic differentiation and geographic distance indicated that gene flow played a more important role in structuring non-Alaskan populations, whereas drift played a more important role in structuring Alaskan populations (R(ST)'s for non-Alaskan populations 0.029 ± 0.007 and 0.083 ± 0.012 for Alaskan populations). Microsatellites also substantiate the bidirectional patterns of gene flow previously uncovered using chloroplast DNA but indicate much greater movement and mixing. Results from our Bayesian analyses also suggest the existence of additional cryptic refugia. However, the locations have been obscured by high gene flow (R(ST) averaging 0.057 ± 0.004).