Patterns of acoustic variation in Cicada barbara Stål (Hemiptera, Cicadoidea) from the Iberian Peninsula and Morocco

Field recordings of the calling song and of an amplitude modulated signal produced by males of Cicada barbara from North Africa and the Iberian Peninsula were analysed in order to assess the geographical acoustic variation and the potential usefulness of acoustic data in the discrimination of subspecies and populations. Sound recordings were digitized and the frequency and temporal properties of the calls of each cicada were analysed. In all regions studied, peak frequency, quartiles 25, 50 and 75% and syllable rate showed low coefficients of variation suggesting inherent static properties. All frequency variables were correlated with the latitude, decreasing from south to north. In addition, most acoustic variables of the calling song showed significant differences between regions, and PCA and DFA analyses supported a partitioning within this species between Iberian Peninsula+Ceuta and Morocco, corroborating mtDNA data on the same species. Therefore, the subspecific division of C. barbara into C. barbara barbara from Morocco and C. barbara lusitanica from Portugal, Spain and Ceuta finds support from the present acoustic analyses, a result which is also reinforced by molecular markers.     

Population structure of Cicada barbara Stål (Hemiptera, Cicadoidea) from the Iberian Peninsula and Morocco based on mitochondrial DNA analysis

We assess the genetic history and population structure of Cicada barbara in Morocco and the Iberian Peninsula, based on analysis of the mitochondrial cytochrome b gene. The divergence between Morocco and the Iberian Peninsula populations was strongly corroborated by the molecular data, suggesting genetically isolated populations with a low level of gene flow. The Ceuta population from Spanish North Africa was more similar to the Iberian populations than the surrounding Moroccan populations, suggesting that the Strait of Gibraltar has not been acting as a strict barrier to dispersal while the Rif Mountains have. The Iberian Peninsula specimens showed a signature of demographic expansion before that which occurred in Morocco, but some of the assumptions related to the demographic parameters should be considered with caution due to the small genetic variation found. The high haplotype diversity found in Morocco implies higher demographic stability than in the Iberian Peninsula populations. These results do not, however, suggest a Moroccan origin for Iberian cicadas; but the most northwest region in Africa, such as Ceuta, might have acted as a southern refuge for Iberian cicadas during the most severe climatic conditions, from where they could expand north when climate improved. The separation of two subspecies within C. barbara (C. barbara lusitanica and C. barbara barbara) finds support with these results.     

Biogeographical connections between the Maghreb and the Mediterranean peninsulas of southern Europe

The glacial–interglacial cycles have caused severe range modifications of species' distributions. In Europe, thermophilic species had to retreat into geographically distinct southern refugia during glaciations. This process produced strong genetic imprints, which are still detectable by the present pattern of genetic differentiation and the distribution of regional diversity. To reveal the biogeographical imprints in the western Mediterranean, we analysed 26 populations of the butterfly Maniola jurtina spread over large areas of its European and North African distribution range. The samples were analysed using allozyme electrophoresis. We detected three genetic groups, divided into Western Europe, Central/Eastern Europe, and Italy with the Maghreb. The North African samples randomly cluster within the Italian samples. Even the population sampled in Morocco is genetically closely related to these samples and not to the geographically neighbouring Iberian ones. Parameters of genetic diversity showed similar values over the whole study area. The observed genetic pattern reflects possible glacial refugia in Europe located in the Iberian Peninsula and the Balkans. For North Africa and Italy, our data reveal a colonization of Africa originating from Italy.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 693–703.     

Phylogeography and conservation genetics of endangered European Margaritiferidae (Bivalvia: Unionoidea)

Margaritifera margaritifera and M. auricularia are among the most endangered freshwater mussels in the world, and the only species of the genus found in Europe. Our genetic study explores allozymic variability (27 loci) and differentiation at the mitochondrial sequence level (partial COI and 16S rRNA gene sequences). The Spanish M. auricularia population showed genetic parameters of variation that were of the same order as those of other freshwater molluscs (though at the lower end of the range), probably permitting its potential recovery. The difference between this species and M. margaritifera was clearly established (ten diagnostic allozymic loci, Nei = 0.462, and mean nucleotide divergence around 9.4%). The M. margaritifera populations analysed showed a certain degree of population genetic structure (according to allozyme data) that was not, however, related to a geographical cline. Nevertheless, two mitochondrial lineages (albeit very closely related) were identified: a northern lineage extending from Ireland to the Kola Peninsula including the western Atlantic coast, and a second cluster distributed from Ireland to the Iberian Peninsula. The phylogenetic relationships between these two species and other related taxa were established. The putative M. m. durrovensis could be considered an 'ecophenotype'. Palaeobiogeographical scenarios are presented and indicate unexpected 'recent' gene flow between M. margaritifera populations that were theoretically isolated in the early Tertiary.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society 2003, 78, 235–252.     

Northern and Southern expansions of Atlantic brown trout (Salmo trutta) populations during the Pleistocene

The phylogeography of Atlantic brown trout ( Salmo trutta ) was analysed using mitochondrial DNA control region complete sequences of 774 individuals from 57 locations. Additionally, the available haplotype information from 100 published populations was incorporated in the analysis. Combined information from nested clade analysis, haplotype trees, mismatch distributions, and coalescent simulations was used to characterize population groups in the Atlantic basin. A major clade involved haplotypes assigned to the Atlantic (AT) lineage, but another major clade should be considered as a distinct endemic lineage restricted to the Iberian Peninsula. The phylogeography of the Atlantic populations showed the mixed distribution of several Atlantic clades in glaciated areas of Northern Europe, whereas diverged haplotypes dominated the coastal Iberian rivers. Populations inhabiting the Atlantic rivers of southern France apparently contributed to postglacial colonization of northern basins, but also comprised the source of southern expansions during the Pleistocene.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 904–917.     

Effects of habitat heterogeneity and local adaptation on the body condition of a forest passerine at the edge of its distributional range

We studied fluctuating asymmetry and feather growth rates as indicators of fitness of blackcaps ( Sylvia atricapilla ) breeding at the border of their distribution range in the Iberian Peninsula. Iberian blackcaps increase their abundance with increasing rainfall and ground cover of brambles. In border habitats, they are sedentary and morphologically different from migrants, suggesting that they could be adapted to peripheral conditions. We tested whether juvenile body condition depends on (1) distance from the centre of the range, (2) mean precipitation or (3) bramble cover. Controlling for environmental variation, we tested for differences between migratory and sedentary populations. Body condition varied across the Iberian gradient in parallel with changes in precipitation. Controlling for this effect, sedentary populations were in better body condition than migratory populations. Our results support the idea that environmental heterogeneity causes fitness to fluctuate across species' ranges, and also that local adaptation may mean that peripheral populations are more than a 'tail end' of the species.  © 2003 The Linnean Society of London . Biological Journal of the Linnean Society , 2003, 78 , 479–488.     

Infraspecific variation and phylogeography of the high-polyploid Iberian endemic Anthoxanthum amarum Brot. (Poaceae; Pooideae) assessed by random amplified polymorphic DNA markers (RAPDs) and morphology

High morphological and ecological diversity has been observed in the north-west Iberian Peninsula endemic Anthoxanthum amarum Brot., for which two different morphotypes (northern and southern) have been described on the basis of qualitative traits and geographical origin. In the present study, a combined molecular and morphological method was applied to ten populations of this species with the following aims: (1) to assess whether the variation observed was taxonomically meaningful; (2) to assess the influence of the environment on the variation in the morphological characters; and (3) to track the potential phylogeographical information deduced from our random amplified polymorphic DNA marker (RAPD) data in order to draw inferences about the past history of this species in the north-west Iberian Peninsula. To achieve these aims, 26 macromorphological characters were recorded in 279 specimens, and 77 RAPD phenotypes were identified in the 79 plants studied. The association analyses performed using the morphological and molecular data showed that no clear separation existed between the morphs, and a strong correlation between qualitative characters and the environment was detected. Moreover, both the multivariate analyses and the assignment test based on RAPD data revealed that the genetic variation was hierarchically structured, and three genetically distinct groups could be identified. Two of these clusters might correspond to different expansion routes proposed in the literature for different plant species in the north-west Iberian Peninsula.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 179–192.     

Taxonomic revision of the genus Succisella (Dipsacaceae) in the Iberian Peninsula

A revision of the three endemic Iberian species of Succisella G. Beck ( S. carvalhoana , S. microcephala and S. andreae-molinae ), based on herbarium studies, SEM photographs and field observations, is presented utilizing morphological, palynological, karyological, biogeographical and ecological characters. The distribution of the species in the Iberian Peninsula is shown in a grid map. Full synonymy is given for all taxa.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 351–364.     

Comparing patterns of nuclear and mitochondrial divergence in a cryptic species complex: the case of Iberian and North African wall lizards (Podarcis, Lacertidae)

Combining different sources of information is essential for a complete understanding of the process of genetic differentiation between species. The Iberian and North African wall lizard ( Podarcis ) species complex has been the object of several studies regarding morphological and mitochondrial DNA variation but, so far, no large-scale survey of nuclear variation within this group has been accomplished. In this study, ten polymorphic allozyme loci were studied in 569 individuals collected across the Iberian Peninsula and North Africa. The obtained data were analysed using both conventional population genetic tools and recent Bayesian model-based clustering methods. Our results show that there are several well-differentiated entities corroborating the major splits observed in mtDNA analyses. These groups correspond not only to the fully recognized species Podarcis bocagei , Podarcis carbonelli , and Podarcis vaucheri but also to multiple forms within the polytypic Podarcis hispanica , all of which have a similar level of differentiation to that observed between the acknowledged species. However, relationships between forms are weakly supported both by population and individual clustering methods, suggesting a scenario of a rapid diversification that contrasts to the clear bifurcating model assumed from previous mtDNA analyses. Individual multilocus analyses report few individuals misassigned or apparently admixed, some of which are most likely explained by the persistence of high levels of ancestral polymorphism. Other admixed individuals, however, are probably the result of limited levels of gene flow between forms.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 121–133.     

Karyological and taxonomic notes on Ophrys (Orchidoideae, Orchidaceae) from the Iberian Peninsula

Karyological information on Iberian Ophrys species is very limited. This paper provides the haploid and diploid chromosome numbers of 11 taxa of sect. Pseudophrys and sect. Ophrys , both of which are well represented in the Iberian Peninsula, and two taxa from Tunisia. The first data on chromosome numbers for O. vasconica (2 n  = 72, 74), O. ficalhoana (2 n  = 36), O. picta (2 n  = 36), O. sphegifera ( n  = 18, 2 n  = 36, 38) and O. passionis (2 n  = 36) are also presented, confirming the stability of the chromosome number in Ophrys . In addition, populations of the group O. omegaifera ( O. dyris and O. vasconica ), together with tetraploidy, pentaploidy and the existence of aneuploid phenomena, are reported for the first time in Iberia. The basic diploid number is always 2 n  = 36. The karyotypes of several species were analysed. Evolutionary trends in Ophrys chromosomes are discussed. Taxonomic and phytogeographical details are provided on several species or groups of species from Iberia.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003 , 142 , 395−406.     

Reproductive ecology of Rhododendron ponticum (Ericaceae) in relict Mediterranean populations

In the southern Iberian Peninsula, Rhododendron ponticum occurs in restricted and vulnerable populations as a Tertiary relict. Population structure and the main phases of the reproductive process were examined in order to shed light on recruitment patterns and limitations. Rhododendron ponticum flowers are self-compatible and attract a diverse array of insects, which are responsible for a considerable number of seeds set in the populations. Nevertheless, only adults form populations, whilst seedlings are scarce and saplings virtually absent (only two juveniles out of 2489 adults sampled). Non-specialized vegetative multiplication by layering was observed. Recruitment failure seems to depend on the scarcity of safe microsites, which are free from drought, for seedling establishment. The observations contrast with R. ponticum 's reputation as an aggressive invader in temperate Atlantic areas. It is proposed that the species shows a variable balance between sexual reproduction and vegetative multiplication depending on environmental conditions. At present, only the latter seems to be prevailing in relict populations in the Iberian Peninsula. This flexible reproductive strategy is also discussed as a mechanism allowing persistence during geological climatic oscillations.  © The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 297–311.     

Two new species of Galium (Rubiaceae) from the Iberian Peninsula

Two new species of the genus Galium are described from the Iberian Peninsula: G. moralesianum Ortega-Olivencia & Devesa, and G. talaveranum Ortega-Olivencia & Devesa. The first grows on limestone or dolomitic-limestone outcrops in south-east Spain (the Segura and Gádor mountain ranges) and shows morphological similarities with G. boissieranum Ehrend. & Krendl, which is endemic to southern Spain (mountains of Málaga). The second inhabits the margins of water courses, alder stands and cork-oak dehesas (parkland-type systems) of the west of the Iberian Peninsula, and is morphologically reminiscent of G. mollugo L. The chromosome numbers of these two species and of G. boissieranum are given.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 177−187.     

Relationships between the close congeners Anthoxanthum odoratum and A. alpinum (Poaceae, Pooideae) assessed by morphological and molecular methods

Anthoxanthum alpinum Löve & Löve has been described as a diploid perennial distributed in northern Eurasia and the high mountains of central and eastern Europe. Difficulties in finding reliable morphological differences between this taxon and the widespread tetraploid Anthoxanthum odoratum L. have resulted in taxonomists treating them as conspecific, despite the cytological differentiation. The purpose of this study was to use different approaches to assess the relationships between close congeners, such as the pair A. odoratum / A. alpinum . Macromorphological, micromorphological, and molecular data were gathered and analysed for 14 populations representing both taxa from Scandinavia and the Iberian Peninsula. Different cluster analyses were performed to study the relatedness between individuals and populations. Subsequently, a principal components analysis was computed on the basis of macromorphological quantitative traits, and principal coordinates analysis was used to analyse qualitative, micromorphological, and random amplification of polymorphic DNA (RAPD) data. An analysis of molecular variance was applied to the molecular data, and the genetic differentiation between samples was measured using the F _ST estimator. The results showed that the geographical origin was more important than the ploidy level in explaining the relatedness between specimens and populations. Moreover, a strong correlation was found between the micromorphological traits and environmental parameters. The results of the analyses do not support the assignment of a specific taxonomic rank to A. alpinum .  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 156 , 237–252.     

Chloroplast DNA variation in the shrub Justicia areysiana (Acanthaceae) endemic to the monsoon affected coastal mountains of the southern Arabian Peninsula

The monsoon affected mountains of the southern Arabian Peninsula harbour in climatically favoured refugia vegetation elements of palaeo-African origin. To understand better the temporal and spatial differentiation of these refugia, chloroplast variation in Justicia areysiana Deflers (Acanthaceae), a shrub species endemic to the Yemeni and Omani mountains close to the Arabian Sea, was studied using PCR-RFLP and chloroplast microsatellite diversity. Eleven haplotypes were characterized and show a distinct geographical distribution pattern with a deep split between populations from south Yemeni fog oases and those from Hawf Mountains/Dhofar region in east Yemen and south Oman. Very limited haplotype diversity within populations (h_S = 0.15) and a high level of population differentiation (G_ST = 0.81) demonstrate the strong genetic isolation of populations from each other. Past oscillations between humid and arid periods connected with glacial and interglacial episodes in the Pleistocene and Holocene are considered responsible for the observed patterns of genetic variation.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 437–444.     

Low population differentiation and high genetic diversity in the invasive species Carduus acanthoides L. (Asteraceae) within its native range in the Czech Republic

Colonizing species are predicted to suffer from reductions in genetic diversity during founding events. Although there is no unique mode of reproduction that is characteristic of successful plant colonizers, many of them are predominantly self-fertilizing or apomictic species, and almost all outcrossing colonizers are self-compatible. Carduus acanthoides comprises a species of disturbed habitats with wind-dispersed seeds that colonizes open spaces of various sizes. Population genetic diversity was expressed by assessing patterns of variation at nine putatively neutral allozyme loci within and among 20 natural populations in its native distribution range in the Czech Republic. Overall, C. acanthoides displayed high levels of genetic diversity compared to other herbaceous plants. The percentage of polymorphic loci was 84.5, with values of 2.37, 0.330, and 0.364 for the mean number of alleles per polymorphic locus ( A ), observed heterozygosity ( H _o), and expected heterozygosity ( H _e), respectively. There was only weak evidence of inbreeding within populations ( f  = 0.097) and very low genetic differentiation among populations ( θ  = 0.085). Analyses of the data provide strong evidence for isolation-by-distance for the whole study area. Even the colonizing species, C. acanthoides , currently supports a substantial amount of allozyme variation at both the species and population levels.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 596–607.     

Taxonomic relevance of the gametophytic generation in a strictly rupicolous fern group: Asplenium seelosii s.l.

The systematics of the Asplenium seelosii complex has been debated for a long time. This complex includes strictly rupicolous plants that live on limestone cliffs mainly in the mountains of south-west Europe: the Alps, the Pyrenees, and several mountain ranges of the eastern Iberian Peninsula. The disjunct distribution of its populations and several morphological characters, i.e. leaf indumentum and the structure of the perispore, have been used to distinguish two species and up to four subspecies. The goal of this study was to determine whether the characters of the gametophytic generation and young sporophytes can contribute to the recognition of the proposed taxa. The analysis of all of these characters together differentiated two groups that correspond to the established species, A. seelosii and A. celtibericum , but not enough differences were found to separate the subspecies in each of them.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 375–386.     

Spatial expansion and population structure of the neotropical malaria vector, Anopheles darlingi (Diptera: Culicidae)

Extensive population structuring is known to occur in Anopheles darlingi , the primary malaria vector of the Neotropics. We analysed the phylogeographic structure of the species using the mitochondrial cytochrome oxidase I marker. Diversity is divided into six main population groups in South America: Colombia, central Amazonia, southern Brazil, south-eastern Brazil, and two groups in north-east Brazil. The ancestral distribution of the taxon is hypothesized to be central Amazonia, and there is evidence of expansion from this region during the late Pleistocene. The expansion was not a homogeneous front, however, with at least four subgroups being formed due to geographic barriers. As the species spread, populations became isolated from each other by the Amazon River and the coastal mountain ranges of south-eastern Brazil and the Andes. Analyses incorporating distances around these barriers suggest that the entire South American range of An. darlingi is at mutation–dispersal–drift equilibrium. Because the species is distributed throughout such a broad area, the limited dispersal across some landscape types promotes differentiation between otherwise proximate populations. Moreover, samples from the An. darlingi holotype location in Rio de Janeiro State are substantially derived from all other populations, implying that there may be additional genetic differences of epidemiological relevance. The results obtained contribute to our understanding of gene flow in this species and allow the formulation of human mosquito health protocols in light of the potential population differences in vector capacity or tolerance to control strategies.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 854–866.     

Volatile leaf oil diversity in the narrow range endemic Eucalyptus argutifolia (Myrtaceae) and its widespread congener Eucalyptus obtusiflora

Variation in the qualitative composition of volatile leaf oil was examined in the rare Eucalyptus argutifolia and its widespread congener Eucalyptus obtusiflora . The results revealed that, consistent with the pattern seen with allozymes, E. argutifolia had less variation within populations than E. obtusiflora . Total leaf oil diversity was also significantly lower in the rare species. As found with allozymes, most leaf oil diversity was within populations, but there was also a significant proportion of the variation between populations (25.2% and 27.3% for E. argutifolia and E. obtusiflora , respectively). There were significant associations between phenotypic distance based on leaf oils and geographical distance and between phenotypic and genetic distance across all populations, but these associations were not evident within species. Factors leading to reduced variation in E. argutifolia appear to affect all types of variation, but the relationships between different types of variation within the species are less apparent.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 738–745.