Habitat‐driven diversification, hybridization and cryptic diversity in the Fork‐tailed Drongo (Passeriformes: Dicruridae: Dicrurus adsimilis)

Despite many ecological and evolutionary studies, the history of several species complexes within the freshwater crustacean genus Daphnia (Branchiopoda, Anomopoda) is poorly understood. In particular, the Daphnia longispina group, comprising several large-lake species, is characterized by pronounced phenotypic plasticity, many hybridizing species and backcrossing. We studied clonal assemblages from lakes and ponds comprising daphnids from several species complexes. In order to reveal patterns of reticulate evolution and introgression among species, we analysed three data sets and compared nuclear, mtDNA and morphological divergence using animals from 158 newly established clonal cultures. By examining 15 nuclear and 11 mitochondrial (12S/16S rDNA) genetic characters (allozymes/restriction enzymes), and 48 morphological traits, we found high clonal diversity and discontinuities in genotypic and morphological space which allowed us to group clones by cytonuclear differentiation into seven units (outgroup D. pulex). In contrast to six groups emerging from nuclear divergence (related to three traditional species, D. cucullata, D. galeata, D. hyalina and three pairwise intermediate hybrids), a seventh group of clones was clearly resolved by morphological divergence: distinct mtDNA haplotypes within one nuclear defined cluster, ‘D. hyalina’, resembled traditional D. hyalina and D. rosea phenotypes, respectively. In other nuclear defined clusters, association between mtDNA haplotype and morphology was low, despite hybridization being bidirectional (reciprocal crosses). Morphological divergence was greatest between young sister species which are separated on the lake/pond level, suggesting a significant role for divergent selection during speciation along with habitat shifts. Phylogenetic analyses were restricted to four cytonuclear groups of clones related to species. mtDNA and nuclear phylogenies were consistent in low genetic divergence and monophyly of D. hyalina and D. rosea. Incongruent patterns of phylogenies and different levels of genetic differentiation between traditional species suggest reticulate evolutionary processes.

A study was conducted to investigate the genetic diversity of C. radicicola from different regions of Oman and its relationship to populations of C. radicicola from other parts in the Arabian peninsula and the USA. A total of 32 C. radicicola isolates from Oman (23), the United Arab Emirates (UAE) (2), Kuwait (1) and the USA (6) were analyzed using sequences of the internal transcribed spacer region of the ribosomal DNA (ITS rDNA) and amplified fragment length polymorphism (AFLP). AFLP analysis of the 32 isolates produced 32 different genotypes and 999 polymorphic alleles (100% polymorphism). The Omani population of C. radicicola was found to be more genetically diverse (H = 0.274) than the USA population (H = 0.186), which may suggest a longer history of C. radicicola in Oman. UPGMA analysis based on Nei's genetic distance as well as phylogenetic analysis based on ITS rDNA sequences separated the USA isolates from those from Oman, UAE and Kuwait with a very high bootstrap support. This was supported by low to moderate levels of genetic differentiation between populations of C. radicicola within the Arabian peninsula (FST = - 0.0297-0.1341) as compared with high levels of genetic differentiation (FST = 0.2939-0.3913) of the population from the USA. These findings may imply that populations of C. radicicola from the USA and the Arabian peninsula have been evolving independently for a considerably long period of time as opposite to constant gene flow between regions in the Arabian peninsula. Analysis of molecular variance (AMOVA) showed that genetic variation is more associated with geographical regions (FST = 0.1807, P < 0.0001) than hosts (FST = 0.0307, P = 0.1681), which may imply the lack of host "cultivar" specialization in C. radicicola.

Bryonia alba (Cucurbitaceae) is a Eurasian herbaceous vine that spreads vegetatively through the production of many stems from a large, tuberous root. The only known U.S. populations of this aggressive apomict are in Idaho, Montana, Utah, and Washington and likely stem from deliberate (and subsequent accidental) introductions. To assess levels and patterns of genetic diversity of B. alba across its introduced range, 23 populations were analyzed for allozyme variation using 12 enzyme systems. On average, 14.9% of loci are polymorphic per population (1.19 alleles per locus)—low values compared to other vascular plant taxa. Mean percent polymorphic loci differed among regions, with the highest values in the Washington and northern Idaho region (20.2%) and Montana (19.0%) and lower values in populations from the Utah and southern Idaho region (7.4%). Observed heterozygosity exceeded that expected at Hardy‐Weinberg equilibrium in six of 23 populations, and a statistically significant excess of heterozygosity was detected at Pgm‐1 in 18 of 23 populations. The level of population differentiation is high (GST = 0.544); however, the level of differentiation among populations within regions is much lower. These results are consistent with the genetic variation and structure expected for an apomict. Based on the level of genetic differentiation among populations, the current disjunct distribution of B. alba in its new range results from two, and possibly three, separate introductions in the western United States. These introductions may stem, in part, from the vine's 19th century use as a medicinal and ornamental plant.

Afzelia xylocarpa is listed as an endangered species on the IUCN World list of Threatened Trees, due to over exploitation for its valuable timber and habitat loss, have resulted in a rapid decline of populations size and local population extinction. Understanding of the processes that determine population genetic structure, gene flow and mating systems is important to conserve and manage the existing populations for this endangered tree species. This study describes the level of genetic diversity and differentiation of fifteen populations of A. xylocarpa in Thailand. Genetic variations at seven nuclear microsatellite loci were examined. The seven nuclear microsatellite loci employed detected a total of 53 alleles (n=432). The nSSRs data indicate that a high level of genetic diversity (HS = 0.575) and low level of genetic differentiation among the 15 examined A. xylocarpa populations. The observed level of genetic differentiation among the 15 populations was low, as revealed by FST = 0.074 and GST = 0.078. The results for the nSSRs suggested that all of the populations in North Eastern, Central Thailand and the Klong Lan populations harbored the high genetic diversity and less divergent from the other populations. Therefore, these populations should be assigned the highest priority for conservation of this species. Key words: Afzelia xylocarpa, genetic diversity, endangered species, tropical tree, conservation.

The evolutionary potential of plant species that reproduce via predominantly clonal means and the conditions under which clonality is favoured are not well known. Long-term clonal reproduction is expected to result in a number of readily detectable genetic signals not present in populations that reproduce by sexual means. We use a hierarchical sampling strategy to assess genotype probabilities and confirm that two rare sister taxa of Banksia ionthocarpa have contrasting modes of reproduction. Banksia ionthocarpa subsp. chrysophoenix reproduces clonally. Populations had low levels of genotypic diversity and were comprised of large clonal patches consisting of many ramets that covered hundreds of square metres and showed little intermixing. The taxon was genetically depauperate (mean Na = 1.97, mean P = 0.66, mean He = 0.282), had high levels of genetic differentiation between populations (θ = 0.481), and populations exhibited excess heterozygosity and linkage disequilibrium (LD) among loci, suggesting historically high levels of clonality. In contrast, the sister taxon B. ionthocarpa subsp. ionthocarpa, which occurs in an area with more than twice the annual rainfall and less extreme minimum and maximum temperatures, showed no evidence of clonality, high levels of genotypic diversity, greater genetic diversity (mean Na = 3.31, mean P = 0.81, mean He = 0.405), lower levels of genetic differentiation between populations (θ = 0.253) and no evidence of excess heterozygosity or LD among loci. We suggest that the development of clonality in subsp. chrysophoenix is associated with its more marginal environment and enhanced by response to recurrent fires.

Populations of Yellowstone cutthroat troutOncorhynchus clarkii bouvieriihave declined throughout their native range as a result of habitat fragmentation, overharvest, and introductions of nonnative trout that have hybridized with or displaced native populations. The degree to which these factors have impacted the current genetic population structure of Yellowstone cutthroat trout populations is of primary interest for their conservation. In this study, we examined the genetic diversity and genetic population structure of Yellowstone cutthroat trout in Idaho and Nevada with data from six polymorphic microsatellite loci. A total of 1,392 samples were analyzed from 45 sample locations throughout 11 major river drainages. We found that levels of genetic diversity and genetic differentiation varied extensively. The Salt River drainage, which is representative of the least impacted migration corridors in Idaho, had the highest levels of genetic diversity and low levels of genetic differentiation. High levels of genetic differentiation were observed at similar or smaller geographic scales in the Portneuf River, Raft River, and Teton River drainages, which are more altered by anthropogenic disturbances. Results suggested that Yellowstone cutthroat trout are naturally structured at the major river drainage level but that habitat fragmentation has altered this structuring. Connectivity should be restored via habitat restoration whenever possible to minimize losses in genetic diversity and to preserve historical processes of gene flow, life history variation, and metapopulation dynamics. However, alternative strategies for management and conservation should also be considered in areas where there is a strong likelihood of nonnative invasions or extensive habitat fragmentation that cannot be easily ameliorated.

BackgroundCoccidioides immitis and C. posadasii cause coccidioidomycosis, a disease that is endemic to North and South America, but for Central America, the incidence of coccidioidomycosis has not been clearly established. Several studies suggest genetic variability in these fungi; however, little definitive information has been discovered about the variability of Coccidioides fungi in Mexico (MX) and Argentina (AR). Thus, the goals for this work were to study 32 Coccidioides spp. isolates from MX and AR, identify the species of these Coccidioides spp. isolates, analyse their phenotypic variability, examine their genetic variability and investigate the Coccidioides reproductive system and its level of genetic differentiation.MethodsCoccidioides spp. isolates from MX and AR were taxonomically identified by phylogenetic inference analysis using partial sequences of the Ag2/PRA gene and their phenotypic characteristics analysed. The genetic variability, reproductive system and level of differentiation were estimated using AFLP markers. The level of genetic variability was assessed measuring the percentage of polymorphic loci, number of effective allele, expected heterocygosity and Index of Association (IA). The degree of genetic differentiation was determined by AMOVA. Genetic similarities among isolates were estimated using Jaccard index. The UPGMA was used to contsruct the corresponding dendrogram. Finally, a network of haplotypes was built to evaluate the genealogical relationships among AFLP haplotypes.ResultsAll isolates of Coccidioides spp. from MX and AR were identified as C. posadasii. No phenotypic variability was observed among the C. posadasii isolates from MX and AR. Analyses of genetic diversity and population structure were conducted using AFLP markers. Different estimators of genetic variability indicated that the C. posadasii isolates from MX and AR had high genetic variability. Furthermore, AMOVA, dendrogram and haplotype network showed a small genetic differentiation among the C. posadasii populations analysed from MX and AR. Additionally, the IA calculated for the isolates suggested that the species has a recombinant reproductive system.ConclusionsNo phenotypic variability was observed among the C. posadasii isolates from MX and AR. The high genetic variability observed in the isolates from MX and AR and the small genetic differentiation observed among the C. posadasii isolates analysed, suggest that this species could be distributed as a single genetic population in Latin America.

Semi-automated fluorescent genotyping of eight polymorphic microsatellite loci was used to assess the level of genetic diversity and population differentiation in Nordic stocks of non-anadromous Arctic charr, Salvelinus alpinus, of both wild and hatchery origin. Highly significant genetic heterogeneity was detected globally across wild and/or hatchery populations and even between populations from lakes separated by as little as six kilometres. The overall level of genetic differentiation among wild populations (FST= 0.360) was substantially higher than that observed between populations of anadromous Arctic charr in Canada using microsatellite data. Cavalli-Sforza and Edward's chord distance was used to construct a neighbour-joining tree and three population clusters were supported with relatively high bootstrap values which included the populations from north-west, north-east and southern Finland respectively. Use of Paetkau et al's individual assignment test further supported the strong differentiation of most populations as well as their classification into the three predicted geographical areas. No significant difference in average allele number or heterozygosity was observed between populations of wild and hatchery origin. However, the effects of hatchery rearing were revealed by a strong increase in the number of deviations from Hardy-Weinberg equilibrium as well as of linkage disequilibrium events in the hatchery stocks compared to natural populations. The population from Lake Saimaa, in south-east Finland, is completely reliant upon aquaculture assistance for its survival. The broodstocks of this population exhibited particularly low levels of genetic variability. Although the hatchery stocks of this population suffer from increased egg and alevin mortality and disease susceptibility, it remains to be determined if this is due directly to a lack of genetic variation as some abundant unstocked natural populations possessed similarly low levels of microsatellite variability.

Despite the theoretical link between the ecology and the population genetics of species, little empirical evidence is available that corroborates the association. Here, I examined genetic variation in 40 co-distributed species of lowland Neotropical rainforest birds that have populations isolated on either side of the Andes, Amazon River, and Madeira River. I found widely varying levels of genetic divergence among these taxa between the same biogeographic barriers. My investigation of the extent to which ecological traits predicted the level of cross-barrier divergence revealed a significant relationship between the forest stratum at which a species forages and the level of within-population and cross-barrier genetic differentiation. Canopy species had statistically lower divergence values across the Andes and two riverine barriers than did understory birds. I hypothesize that the association reflects an effect of dispersal propensity on the geographic structuring of genetic variation, and, consequently, on the ancestral and extant effective population sizes of each species. This is the first large-scale avian comparative study to document a significant association between ecological traits of a species and its level of genetic differentiation. I examined further the contrasting genetic patterns revealed previously by comparing the range-wide mitochondrial (mtDNA) phylogeography of two canopy and two understory species of lowland Neotropical rainforest birds. All species exhibited divergence between cross-Andean populations. Unlike canopy species, understory birds were structured at smaller spatial scales, particularly across riverine barriers of the Amazon basin. Surprisingly, estimates of isolation-by-distance, a proxy for dispersal propensity, are similar within areas of endemism for all taxa suggesting levels of gene flow are comparable through contiguous habitat in canopy and understory species. Lastly, I examined the multilocus phylogeography of three previously studied species with contrasting mtDNA patterns to investigate the role of historical demography in cross-Andean divergence. Demographic estimates using an isolation-with-migration model suggest among-taxa variance in cross-Andean divergences reflects a history of staggered isolation versus a simultaneous isolating event. Nuclear sequence data reveal asymmetrical gene flow in two species marked by relatively shallow cross-Andean divergence, further evidence of differential effectiveness of the Andes as a barrier to gene flow among co-distributed taxa.

Sequence analyses of the non-coding, control region (CR) and coding region of the ND4-tRNA(Ser) genes in the mitochondrial DNA (mtDNA) were conducted for populations of the ayu Plecoglossus altivelis altivelis and the Ryukyu-ayu P. a. ryukyuensis. The level of genetic differentiation between the two subspecies evaluated from the CR data was substantially low, when comparing with that estimated from ND4-tRNA(Ser) gene region data, as well as those from nuclear genome data sets. By contrast, the differentiation between subspecies in the ND4-tRNA(Ser) gene region was substantial, being consistent with the results from the previous nuclear genome analyses. Results of UPGMA and minimum spanning network analyses also implied the unexpected ceiling of genetic differentiation in the CR. These results suggest that the CR does not reflect accurately the level of overall genetic differentiation between the populations of the ayu, but other coding regions of the mtDNA do reflect it so that the mtDNA on the whole may function as a rich source of useful markers for genetic assessment of populations of this species.

Levels of genetic differentiation between populations can be highly variable across the genome, with divergent selection contributing to such heterogeneous genomic divergence. For example, loci under divergent selection and those tightly physically linked to them may exhibit stronger differentiation than neutral regions with weak or no linkage to such loci. Divergent selection can also increase genome-wide neutral differentiation by reducing gene flow (e.g. by causing ecological speciation), thus promoting divergence via the stochastic effects of genetic drift. These consequences of divergent selection are being reported in recently accumulating studies that identify: (i) 'outlier loci' with higher levels of divergence than expected under neutrality, and (ii) a positive association between the degree of adaptive phenotypic divergence and levels of molecular genetic differentiation across population pairs ['isolation by adaptation' (IBA)]. The latter pattern arises because as adaptive divergence increases, gene flow is reduced (thereby promoting drift) and genetic hitchhiking increased. Here, we review and integrate these previously disconnected concepts and literatures. We find that studies generally report 5-10% of loci to be outliers. These selected regions were often dispersed across the genome, commonly exhibited replicated divergence across different population pairs, and could sometimes be associated with specific ecological variables. IBA was not infrequently observed, even at neutral loci putatively unlinked to those under divergent selection. Overall, we conclude that divergent selection makes diverse contributions to heterogeneous genomic divergence. Nonetheless, the number, size, and distribution of genomic regions affected by selection varied substantially among studies, leading us to discuss the potential role of divergent selection in the growth of regions of differentiation (i.e. genomic islands of divergence), a topic in need of future investigation.

The global decline of freshwater mussels and their crucial ecological services highlight the need to understand their phylogeny, phylogeography and patterns of genetic diversity to guide conservation efforts. Such knowledge is urgently needed for Unio crassus, a highly imperilled species originally widespread throughout Europe and southwest Asia. Recent studies have resurrected several species from synonymy based on mitochondrial data, revealing U. crassus to be a complex of cryptic species. To address long-standing taxonomic uncertainties hindering effective conservation, we integrate morphometric, phylogenetic, and phylogeographic analyses to examine species diversity within the U. crassus complex across its entire range. Phylogenetic analyses were performed using cytochrome c oxidase subunit I (815 specimens from 182 populations) and, for selected specimens, whole mitogenome sequences and Anchored Hybrid Enrichment (AHE) data on ∼ 600 nuclear loci. Mito-nuclear discordance was detected, consistent with mitochondrial DNA gene flow between some species during the Pliocene and Pleistocene. Fossil-calibrated phylogenies based on AHE data support a Mediterranean origin for the U. crassus complex in the Early Miocene. The results of our integrative approach support 12 species in the group: the previously recognised Unio bruguierianus, Unio carneus, Unio crassus, Unio damascensis, Unio ionicus, Unio sesirmensis, and Unio tumidiformis, and the reinstatement of five nominal taxa: Unio desectusstat. rev., Unio gontieriistat. rev., Unio mardinensisstat. rev., Unio nanusstat. rev., and Unio vicariusstat. rev. Morphometric analyses of shell contours reveal important morphospace overlaps among these species, highlighting cryptic, but geographically structured, diversity. The distribution, taxonomy, phylogeography, and conservation of each species are succinctly described.

Genetic diversity is not always congruent with phenotypic heterogeneity, resulting in cryptic species complexes which cause a great struggle for scientists trying to define ‘species’ and describe relationships among taxa. Anatololacerta is a lizard genus distributed in southern and western Anatolia and some neighboring Aegean islands. Three morphospecies were recognized in Anatololacerta but a recent molecular study revealed the presence of cryptic diversity within the genus which led to the raise of a subspecies to species level. Currently the genus includes the species A. anatolica, A. danfordi, A. budaki and A. pelasgiana. Using a comprehensive sampling concerning both the number of specimens (218 specimens) and the genetic markers (3 nuclear and 3 mitochondrial), we performed phylogenetic analyses including tree reconstruction, species delimitation and divergence times estimation. The results revealed the occurrence of one more cryptic lineage which should be regarded as a separate species for which the name A. ibrahimi stat. nov. has priority. The existence of five well differentiated species with parapatric distributions in Anatololacerta is strongly supported. There is also evidence of recent and rapid radiation of the genus which probably causes phylogenetic relationships between these species to remain largely unresolved. At last, we proceeded to some nomenclatorial changes: The current name A. budaki was synonymized with A. pelasgiana because specimens of the type-locality of A. budaki are assigned genetically to A. pelasgiana. The genetic lineage including specimens currently assigned to A. budaki was named A. finikensis stat. nov., raising the subspecies A. b. finikensis to species level.

A comparative analysis of the genetic variability and differentiation of rare medicinal ginseng species, Panax vietnamensis Ha et Grushv. and P. ginseng C.A. Meyer, was carried out using inter-simple sequence repeat markers. It was demonstrated that all the genetic diversity parameters of Vietnamese ginseng were high and considerably exceeded those of P. ginseng. On the contrary, the level of genetic differentiation was higher in true ginseng. It is suggested that the differences in the levels of genetic variability and differentiation of the two ginseng species were influenced by the demographic history, peculiarities of the reproductive system, and human activity.

Distribution margins constitute areas particularly prone to random and/or adaptive intraspecific differentiation in plants. This trend may be particularly marked in species discontinuously distributed across mountain ranges, where sharp geographic isolation gradients and habitat boundaries will enhance genetic isolation among populations. In this study, we analysed the level of neutral genetic differentiation among populations of the long-lived shrub Daphne laureola (Thymelaeaceae) across the Baetic Ranges, a glacial refugium and biodiversity hotspot in the western Mediterranean Basin. Within this area, core and marginal populations of D. laureola were compared with regard to their spatial isolation, size, genetic diversity and differentiation. A spatially explicit analysis conducted on the vast majority of the species' known populations in the study area (N = 111) showed that marginal populations (western and eastern) present larger spatial isolation than core populations, but are not smaller. We compared genetic diversity and differentiation between core and marginal populations using a subsample of 15 populations and 225 amplified fragment length polymorphism (AFLP) markers. Core and marginal populations did not differ in genetic diversity, probably because of the occurrence of large populations on the local margins. Western populations were strongly differentiated from the other populations. In addition, spatial and genetic differentiation among populations was larger on the western margin. Eastern populations constituted a genetically homogeneous group closely related to core populations, despite their greater spatial isolation. Results suggest that studies on phenotypic differentiation between core and marginal populations of D. laureola, and presumably other species having discontinuous distributions across the Baetic ranges, should take into account geographical differences in levels of genetic differentiation between the different distribution borders.